KDM1A inhibitors for the treatment of disease

ABSTRACT

Disclosed herein are new compounds and compositions and their application as pharmaceuticals for the treatment of diseases. Methods of inhibition of KDM1A, methods of increasing gamma globin gene expression, and methods to induce differentiation of cancer cells in a human or animal subject are also provided for the treatment of diseases such as acute myelogenous leukemia.

This application claims the benefit of priority of U.S. ProvisionalApplication No. 62/115,474, filed Feb. 12, 2015, the disclosure of whichis hereby incorporated by reference as if written herein in itsentirety.

The present disclosure relates to new compounds and compositions andtheir application as pharmaceuticals for the treatment of diseases.

Inhibiting the enzyme KDM1A (also known as lysine-specific demethylase1, LSD1, Flavin-containing Amine Oxidase Domain-Containing Protein,AOF2, BRAF35-HDAC Complex Protein BHC110, FAD-Binding ProteinBRAF35-HDAC Complex), may alter gene expression in cells sufficient torestore their proper physiologic function or that of the tissue, organor the patient as a whole. This may be achieved either by enhancingtranscription of a gene or genes that are pathologically silenced, e.g.,as is the case in some cancer cells and heritable diseases, ordecreasing transcription of a gene or genes participating in thepathological state. As such, inhibiting KDM1A would be useful for thetreatment of diseases such as cancer and heritable diseases such asWilson disease, cardiomyopathies, and hemoglobinopathies.

Gene expression is regulated through the recruitment of the RNApolymerase II transcription apparatus to the DNA template. Theprobability of this large multi-protein complex arriving near or at thestart of DNA transcription and progressing through the entire codingregion of a gene is determined in part by specific DNA sequences calledpromoters and enhancers, modifications of DNA sequence in the vicinityof the start of transcription, proteins bound to DNA and the topology ofthe DNA template itself. Factors enhancing the probability of RNAsynthesis of protein-coding genes are known as transcription factorssome of which participate in the transcription of all protein-codinggenes and some of which are specific for the transcription of individualgenes.

One major mechanism of transcription control consists of limiting thephysical accessibility of the transcriptional regulatory regions toproteins that can activate or complete transcription; proteins bound topromoter or enhancer DNA sequences can occlude activating factors frombinding to these DNA sequences resulting in fewer transcriptioninitiations or extension of the activated progressing RNA polymerasecomplex. Likewise, topological constraints that do not allow thetemplate DNA to unwind sufficiently to permit the steady progression ofRNA polymerase on the template also serve to limit transcription rates.

The most important general factors influencing RNA synthesis using a DNAtemplate in vivo are modifications of histones proteins that controlamong other factors the topology of the DNA template for transcriptionand its accessibility by the RNA polymerase complex. A small family ofhistone proteins—H2A, H2B, H3 and H4—combines to create a scaffoldcalled the histone octamer upon which DNA is spatially and topologicallyorganized into a regular repetitive structure called the nucleosomealong the length of DNA. The conglomerate of histones, other proteins,various RNAs and DNA is called chromatin. Both DNA and histones arechemically modified in such a way as to attract and bind or repel otherproteins with the effect of enhancing or repressing transcription.

The modification of DNA and associated RNAs and proteins that influencethe regulation of transcription and replication that does not involvesubstitution of the canonical DNA bases is termed epigenetic. Theseepigenetic influences involve reversible chemical modifications of thefour DNA bases themselves or post-translational chemical changes to thechromatin proteins and RNDs that associate with DNA. These epigeneticprocesses can play a pivotal role in activating or silencing theexpression of a gene; in addition, the epigenetic modifications can bemaintained for the life of an organism or can be dynamically modified inresponse to specific biochemical signals that originate eitherinternally within the cell or extracellularly. These chromatinalterations can happen quickly or be very stable, e.g., during thehormonal induction of gene expression, chromatin structure at a specificlocus can change radically within seconds to permit maximaltranscription or chromatin structure can be modified to fully suppressgene expression, a state of chromatin which can be stably maintainedover multiple cell divisions and even transgenerationally.

The methylation of cytosine at the 5′ position is a common DNA basemodification that is in turn recognized by a class of proteins mostoften associated with transcriptional repression. Similarly, histoneproteins are chemically modified but with a wider variety of chemicaladducts each of which either alone or in combination enhances orrepresses transcription of nearby genes. These histone modificationsinclude, among others methylation, acetylation, sumoylation,phosphorylation, ubiquitylation, and myristoylation are recognized byother chromatin-associated proteins that in turn influence transcriptionrates and DNA replication. The dynamic state of gene expression and theassociated chromatin states imply that histone modifications are notpermanent but instead are added and removed according to the needs ofthe cell for specific gene products at specific times during ontogeny,adult life and the changing influences of the environment. Indeed, thespecific chemical modifications of histones are each made by classes ofenzymes acting at specific sites. These histone-modifying enzymes are inturn subject to tight regulation. These enzymes can potentially betargeted by compounds that inhibit their activity with the consequenceof altering gene expression in a therapeutic manner.

Changes in the state of histone methylation are now known to playcritical roles in normal regulation of the cell cycle and growth, theresponse to DNA damage and stress, and pre-natal development includingdifferentiation. Pathological states such as cancer are associated withaltered patterns of histone modifications and dysregulatedhistone-modifying proteins including chromatin-modifying enzymes. Theneed to closely regulate histone modifications is evidenced by theassociation of histone methylation status with human morbidity includingageing.

Histone methylation can occur on any of the three basic amino acidresidues—lysine (K), arginine (R), and histidine (H). Methylation ofhistone H3 on lysines at positions 4 (H3K4), 9 (H3K9), 27 (H3K27), 36(H3K36) and 79 (H3K79) are among the best studied of histonemodifications that influence gene expression. Lysine tri-methylation(Kme3) on histone 3 (H3) at position 4 (H3K4me3) is a histone markgenerally associated with activation of gene expression while H3K9me1 orH3K27me3 are associated with the repression of gene transcription.H3K4me1 is associated with DNA enhancers of gene transcription whileH3K4me3 is associated with gene promoter activity. Likewise, loss of themethyl group at H3K4 is associated with repression of gene expression.Thus, the addition and removal of methyl groups at H3K4 constitutes agene transcription switch. It is also evident that lysine can bemodified with a mono-, di- or tri-methyl groups, each modificationhaving a different biological effect through the attraction of differentproteins recognizing those specific methylation modifications at thatsite.

A critical aspect of the regulation of the state of histone methylationis the recruitment of methyltransferases and demethylases to specificgenetic loci. DNA sequence-specific binding proteins includingtranscription factors are one class of proteins responsible for thisrecruitment through the assemblage of protein complexes that bind thesemethyl-transferring enzymes. A well-studied example is the Drosophilamelanogaster trithrorax group (TrxG) response elements (TREs) whichrecruit the H3K4 methyltransferase, TRX, to specific genes viatranscription factors that recognize the TRE DNA sequence.

The histone methylation marks are recognized by methyl-binding domainsin a diverse group of proteins; these domains include PHD fingers, WD40and ankyrin repeats, CW and PWWP domains, and the Royal superfamily ofproteins. These proteins, in turn, determine which additional activitiesare recruited into chromatin sites and ultimately the state oftranscription at a given locus. Indeed, depending on whichmethyl-recognition protein binds the marked histone, the samemethyl-lysine modification can have opposing effects on transcription.H3K4me2 and H3K4me3 are associated with transcriptional activation, butwhen bound by the PHD-domain-containing co-repressor protein Inhibitorof Growth family member 2 (ING2), an associated histone deacetylasecomplex is stabilized repressing gene expression. Thus, these effectorproteins recognizing the methyl-lysine histone modificationssignificantly influence the level of transcriptional activity.

The ability to alter gene expression selectively by modifying the stateof chromatin allows a novel therapeutic strategy to induce or de-repressthe expression of genes that can provide a benefit, especially for geneswhose expression has been suppressed by pathological mechanism as in thecase of some cancers or suppressed by physiologic mechanism but whode-repression can phenotypically suppress mutations in paralogous geneswith complementary function.

Many genes within a genome are members of gene families as a consequenceof gene duplication. These genes are termed paralogs of one another.Following gene duplication, patterns of expression of two genes willevolve in a distinct manner in part to control the effects of genedosage. Following gene duplication, random genetic drift arising fromnaturally occurring mutations and the subsequent selection of nucleotidesequence is commonly observed first in non-coding regions of duplicatedgenes, often in transcriptional regulatory regions. DNA changes inregulatory sequences can influence any or all aspects of geneexpression: the magnitude of expression, its developmental timing,induction by stimuli outside the cell including hormonal or metabolicsignals, and the cell type in which expression is restricted. Ininstances in which the duplication is recent in evolutionary time orwhere natural selection has maintained a high degree of protein-codingsequence similarity, the gene product of one paralog, gene A, cancomplement the pathological loss or silencing of the other paralog, geneB, if expression of gene A is not limiting in the same cell.

Altering patterns of gene expression may offer profound therapeuticbenefits for genetic conditions in which enhanced expression of aparalogous gene “rescues” a phenotype caused by a mutation in a paralog.This might be called autologous gene complementation. In the case ofWilson disease caused by mutations in ATP7B, enhanced expression bypharmacologic induction of ATP7A, a closely related copper transporterprotein, might rescue mutations in ATP7B, another copper transporter.The basic function of each copper transporter protein has been preservedbut following the duplication of the common ancestral gene, theexpression of these two genes has been separated spatially, one confinedto intestinal enterocytes, the other to hepatocytes. This is one of manyexamples of paralogous gene in which one gene can complement the loss ofthe second if appropriately expressed in the same cell or tissue.

A notable example of a paralogous gene family is the well-studied alphaand beta family of globin genes coding for the alpha and beta subunitsof hemoglobin. Five beta-like genes each arising by gene duplication arearrayed next to each other on chromosome 16 with each gene beingtranscribed in a temporally-specific manner throughout the 9 months ofhuman embryonic and fetal development. The five beta-like globinproteins share a high degree of protein sequence similarity, so much sothat genetic mutations inactivating the adult beta globin gene can beclinically silent if expression of any one of the other 4 subunitmembers of the beta-like globin family is adequate. Activation ofexpression and subsequent transcriptional silencing of each specificembryonic and fetal beta-like globin gene is regulated in part byepigenetic mechanisms. The rescue of mutations in the beta globin gene,mutations which are responsible for diseases such as thalassemia majoror sickle cell anemia, by transcriptional induction of one or more ofthe other beta-like genes through the pharmacologic manipulation ofepigenetic silencing would be clinically beneficial. Autologousactivation with a pharmacologic agent of a functionally complementaryparalog of a mutated or pathologically silenced gene may be a moresuccessful therapeutic strategy than replacing or repairing the mutatedgene with a wild-type (normal) copy.

Interest in influencing the activity of histone modifications fortherapeutic effect derive from observations that the expression ofspecific genes under epigenetic control could be altered by alteringepigenetic marks such as methylation. In the case of cancer, loss ofspecific histone methylation marks concomitant with overexpression ofhistone demethylases is associated with the recurrence of those cancerswith attendant poorer outcomes. These studies suggest that specifictumor suppressor genes are silenced by loss of methylation modificationsthat in turn enhance the survival and growth potential of neoplasticcells. This had led to the proposition that inhibition of histonedemethylase activity might have therapeutic value.

KDM1A (also known as Lysine-Specific Demethylase 1 (LSD1) or AOF2 orBHC110) was the first enzyme with specific lysine demethylase activityto be described demonstrating unequivocally that histone modificationsare reversible rather than permanent. Among its demethylase substrates,KDM1A is a histone H3 lysine demethylase that catalyzes the oxidativedemethylation of H3K4me1 or me2 and H3K9me1 or me2 but not the substrateH3K4me3. The enzyme also demethylates non-histone proteins such as p53and Gfi1. KDM1A contains an amine oxidase domain that demethylates H3Kmesubstrate in a flavin adenine dinucleotide (FAD)-dependent mannersimilar to other monoamine (MAO) and polyamine oxidase inhibitors.Indeed, non-specific inhibitors of MAO enzymes can inhibit thedemethylase activity of KDM1A

KDM1A is over-expressed in many human cancers including Wilm's tumor,small-cell lung, bladder, prostate, breast, head & neck, colon, andovarian cancer and associated with more frequent relapses. KDM1A isrequired for transcriptional regulation mediated by the androgenreceptor in prostate cancer, the estrogen receptor in breast carcinomas,and the TLX receptor in neuroblastoma. Knockdown of KDM1A expressiondecreases proliferation of cancer cells. KDM1A is also overexpressed incancer cells that are nuclear hormone receptor-independent includingER-negative breast. Potent, selective small molecule inhibitors of KDM1Ashould be useful for treatment of these and other cancers in which KDM1Aactivity is overabundant.

The structure and state of chromatin can also influence the ability of apathogenic virus to insert into host DNA, undergo transcription andreplicate. Infection by the alpha herpes viruses herpes simplex virus(HSV) and varicella-zoster virus (VSV) effect the remodeling ofchromatin after infection of host cells to counter the rapid depositionof nucleosomes containing histones with transcriptional repressive marksby employing virus-encoded transcription factors to recruit the hostHCF-1 co-activator complex that contains KDM1A and the histone H3K4methyltransferases Set1 or MLL family members. It has been demonstratedthat inhibition of KDM1A in cells infected with HSV1 inhibits HSV IEgene expression, suppresses lytic infection and reduces viral loads.Similarly, inhibiting KDM1A causes a decrease in the expression of theimmediate early genes in cells infected with human cytomegalovirus andadenovirus suggesting a broader role for KDM1A in viral pathogenesis.

The influence KDM1A activity has on the transcription of specific genesis dependent on recruitment of KDM1A to a specific gene promoter regionvia DNA binding proteins. In the case of androgen-dependent geneexpression, KDM1A associates with the androgen steroid receptor whichspecifically targets DNA binding sites in the promoters ofandrogen-responsive genes. Thus, proteins that bind KDM1A determinewhere along the chromosome the demethylase activity is targeted. Manyproteins have been reported to interact with KDM1A including the CoREST,CtBP, NuRD, BRAF35 complexes, DNMT1, MTA1/2, Mi2beta, RbAp46/48, HDAC1,2, and 3, TIF1beta, Blimp-1, ZNF217 and ZNF198, a subset of which formlarger and in some cases complexes that mutually exclude one another.The KDM1A/CoREST complex which may also include DNMT1 and NuRD amongother factors is particularly important for the repression of expressionof specific genes.

KDM1A is recruited to the promoter region of genes through site-specifictranscription factors. Such factors include among others the androgenreceptor, the estrogen receptor alpha, Snail1, Slug, HIV Tat, ZEB1,RBP-J, PIT1, REST, NR2C1, NR2C2 and isoforms of Gfi1b. Thesetranscription factors can recruit KDM1A to participate in activation ofgene expression or silencing of gene expression depending on the celltype and the specific transcription factors.

Many of the enzyme activities that regulate the state of chromatin areinfluenced allosterically or require as co-factors metabolicintermediates, mediators or end-products of cell metabolism. Theseintermolecular relationships between gene expression and metabolismprovide cells with signaling pathways connecting the external andinternal cellular environment including nutrients with mechanismsmodulating gene expression. This cellular sensing can alter both shortand long term adjustments to gene expression patterns constituting anepigenetic memory of historical metabolic states and environmentalconditions. For example, beta-hydroxybutyrate, a product of long chainfatty acid metabolism and a major source of energy for mammals duringstarvation or prolonged exertion, inhibits class I histone deacetylases(HDAC) but not class 2b HDAC. Thus the effects of starvation andnutrient loss can be epigenetically coded and preserved. Acetyl-coenzymeA, nicotinamide adenine dinucleotide (NAD) and alpha-ketoglutarate alsoinfluence histone methylation and acetylation states.

Flavin adenine dinucleotide (FAD) is a required co-factor for KDM1A.FAD, in conjunction with NAD and NADP act as cellular redox sensors.KDM1A temporarily converts FAD to FADH after which an electron acceptor,likely O₂ and others, completes the catalytic cycle by regenerating FADand H₂O₂. Thus, the cellular redox state influences KDM1A activity bothby its ability to oxidize FADH and other electron acceptors. In ageneral sense, chromatin states, hence gene expression, can be alteredby the variable concentrations of metabolic intermediates and in thespecific case of KDM1A that activity is entirely dependent on FAD whoseconcentration fluctuates as a function of the energetic economy of thecell. In addition, it has been shown that inhibition of KDM1A can lowerserum glucose, reduced hepatic glycogen, and is a powerful insulinsecretagogue. Pharmaceutical manipulation of KDM1A activity may thusprove useful for the treatment of diseases that represent pathologicalaberrations of the energy status of the cell including metabolicsyndrome, dyslipidemias, diabetes, obesity, anorexia, failure to thrive,cachexia, lipodystrophies, and steatohepatitis.

The steroid hormones estradiol and testosterone and related compoundplay a key role in both normal development and in pathological statessuch as breast and prostate cancer in which tumor cell growth isdependent on hormonal signaling. The biological effects of steroidhormones are mediated by structurally and functionally distinctligand-binding receptors that function as a transcription factorrecruited to a specific DNA binding site. The ligand-bound steroidreceptors act as the principal transcriptional regulator of hormoneeffects. Transcriptional activation of gene expression for allsteroid-dependent hormones is dependent on chromatin structure and thepresence of co-factors. The estrogen receptor employs, for example, theco-factors SRC1, SRC2, AIB1, PELP1, CBP, p300, PCAF, CARM1, PRMT1 andco-repressors such as NCoR, SMRT and MTA1. The transcriptional responseto hormone stimulation is dependent on the interaction of theseco-factors and repressors as well as the state of chromatin, especiallymodification of histones by histone-modifying enzymes associated withthe co-regulators. Both estrogenic and androgenic hormone stimulationinduces several histone modifications at the promoters of target genesthat alter the acetylation, phosphorylation and methylation state oflocal histones. To affect the maximal rate of transcription for ahormone-responsive gene, KDM1A activity is required. Thus, KDMA1 shouldprove useful as a therapeutic target of pharmaceuticals in blunting orablating the hormone-dependence of tumor cells. This same therapeuticlogic applies to other ligand-dependent transcription factors whosetranscriptional activation is partly or wholly dependent on KDM1Aactivity to alter chromatin states sufficiently to facilitatetranscription—examples of these would include the vitamin D, retinoidand lipid-activated receptors.

Numerous therapeutic agents have been identified that have the effect ofaltering gene expression acting either directly on proteins, generallyenzymes, that alter chromatin states or indirectly. Though the precisemechanisms of their action have not all been fully elucidated, thosemechanism can be inferred from our understanding of the proteincomplexes that participate in the activation of specific geneexpression. These agents include 5′-azacytidine and 5′-aza-2′deoxycytidine (decitabine) which inhibit DNMT1 or other DNAmethyltransferases known to be present and active at promoter sites ofsilenced genes such as gamma globin promoter; vorinostat andpanobinostat or other inhibitors of histone deacetylase (HDAC) enzymes;hydroxyurea (HU), valproate and sodium butyrate and its analogues eachof which may interfere with the activity of orphan nuclear receptors.All of these agents enjoy some clinical use principally in themanagement of neoplastic disease. Though some clinical utility of theseagents for other disease states has been demonstrated, these agents havenot been widely adopted because of their modest therapeutic effects andtheir toxicity.

The use of agents that inhibit any enzymatic activity resident in theprotein complex bound to gene promoter has the potential to disrupt therepression of gamma globin gene expression and result in increasedlevels of fetal hemoglobin also known as hemoglobin F (HbF). Suchtargets include any of the interfaces of the specific protein-proteincontacts, for example, the NuRD complex and KDM1A; the DNA bindingrecognition domains of, for example, NR2C1 and NR2C2; the ligand bindingdomains of, for example, NR2C1 and NR2C2; the enzyme activities such aslysine demethylase, for example, KDM1A; histone deacetylases (HDAC), forexample HDAC1, 2, or 3; DNA methyltransferases, for example, DNMT1.

There remains a need for compositions and methods for altering geneexpression in cells and tissues sufficient to restore the cell or tissueto normal physiologic function including, e.g., appropriate apoptosis inthe case of cancer, or to alter the pathological phenotype of the cell,tissue, organ or organism by inducing the expression of one or moregenes sufficiently to suppress the pathological state.

Accordingly, the inventors herein disclose new compounds, compositionsand methods for treating diseases associated with KDM1A activity.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. shows an XRPD diffractogram of amorphous Example 1 bis-tosylate.

FIG. 2 shows XRPD diffractograms of Example 1 bis-tosylate Form 2; theupper was recrystallized from semicrystalline solid.

FIG. 3 shows the 1H NMR spectrum of Example 1 bis-tosylate Form 2recrystallized from semicrystalline solid.

FIG. 4 shows the DCS and TGA of Example 1 bis-tosylate Form 2recrystallized from semicrystalline solid.

FIG. 5 shows XRPD diffractograms of Example 1 bis-tosylate Form 2; theupper was synthesized in ACN.

FIG. 6 shows the 1H NMR spectrum of Example 1 bis-tosylate Form 2synthesized in ACN.

FIG. 7 shows the DCS and TGA of Example 1 bis-tosylate Form 2synthesized in ACN.

FIG. 8. shows XRPD diffractograms of Example 1 bis-tosylate forms 2 (A),3 (E), 4 (D), and 5 (B and C, from high-resolution and high-throughputscans, respectively); extra peaks compared to Form 2, noted with dashedlines, prompted additional characterization of Forms 3, 4, and 5.

FIG. 9. shows XRPD diffractograms of Example 1 bis-tosylate form 2 andsemicrystalline form 3 from tetrahydrofuran (F) and toluene (G); extrapeaks compared to Form 2, noted with dashed lines, prompted additionalcharacterization.

FIG. 10 shows XRPD diffractograms of solid material recovered in saltexperiments in various solvents. A is a weakly crystalline sulfate; B isamorphous phosphate; C is amorphous tartrate; D is amorphous sulfate; Eis amorphous phosphate; F is amorphous tartrate; G is amorphousfumarate; H is amorphous sulfate; I is amorphous phosphate; J isamorphous tartrate; and K is weakly crystalline fumarate.

FIG. 11 shows XRPD diffractograms of solid material recovered in saltexperiments in various solvents after maturation. A is amorphoushydrochloride from MTBE; B is partially crystalline sulfate pattern 1; Cis amorphous mesylate from IPA; D is partially crystalline phosphatepattern 1; E is amorphous tartrate; F is amorphous fumarate; G isamorphous sulfate; H is amorphous mesylate from MTBE; I is amorphousphosphate; J is amorphous tartrate; K is amorphous fumarate; L isamorphous sulfate; M is amorphous mesylate from MTBE; N is amorphousphosphate; O is amorphous tartrate; and P is weakly crystalline fumaratepattern 1.

FIG. 12 shows XRPD diffractograms of solid material recovered in saltexperiments in methylethyl ketone. A is amorphous sulfate; B isamorphous oxalate; C is galactaric acid; D is amorphous tartrate; E isamorphous sulfate; F is p-TSA (tosylate) Form 1; G is amorphous oxalate;H is galactaric acid; I is weakly crystalline ascorbate; J is amorphoustartrate.

FIG. 13 shows XRPD diffractograms of solid material recovered in saltexperiments in methylethyl ketone after maturation. A is the sulfatepattern 2; B is the oxalate Form 1; C is galactaric acid; D is amorphoustartrate; E is galactaric acid; and F is weakly crystalline tartrate.

FIG. 14 shows XRPD diffractograms of solid material recovered in saltexperiments in MeCN. A is weakly crystalline sulfate; B is weaklycrystalline oxalate; C is galactaric acid; D is amorphous sulfate; E istosylate Form 2; F is amorphous oxalate; G is galactaric acid; H isamorphous tartrate.

FIG. 15 shows XRPD diffractograms of solid material recovered in saltexperiments in MeCN after maturation. A is sulfate pattern 3; B isoxalate form 1; C is galactaric acid; D is amorphous tartrate; E isamorphous oxalate; F is galactaric acid; G is amorphous tartrate.

FIG. 16 is a high-resolution XRPD diffractogram of the tartrate form 2salt.

FIG. 17 is a high-resolution XRPD diffractogram of the oxalate form 1salt.

FIG. 18 is a high-resolution XRPD diffractogram of the tartrate form 1salt.

FIG. 19 is a high-resolution XRPD diffractogram of the tartrate form 3salt.

FIG. 20 is a high-resolution XRPD diffractogram of the oxalate form 2salt.

FIG. 21 shows XRPD diffractograms of sulfate salt forms from the secondsalt experiment.

FIG. 22 shows XRPD diffractograms of benzenesulfonate salt forms fromthe second salt experiment.

FIG. 23 shows XRPD diffractograms of oxalate salt forms from the secondsalt experiment.

FIG. 24 is an XRPD diffractogram of fumarate salt forms from the secondsalt experiment.

FIG. 25 is an XRPD diffractogram of the besylate salt from the secondsalt experiment.

FIG. 26 is an XRPD diffractogram of L-malate salt from the second saltexperiment.

FIG. 27 is an XRPD diffractogram of L-malate salt from the second saltexperiment.

DETAILED DESCRIPTION

Accordingly, provided herein are compounds of the formula (I):

or a salt, polymorph, or solvate thereof, wherein:

Y is chosen from a bond, NR^(4a), O, C(O)NH, NHC(O), S, SO₂, CHOH, andCH₂;

Z is chosen from a bond, NR^(4b), O, C(O)NH, NHC(O), S, SO₂, and CH₂;

m is chosen from 0, 1, 2, 3, 4, and 5;

n is chosen from 0, 1, 2, and 3;

R¹ and R² are each independently chosen from alkyl, aminoalkyl,alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl,cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl,heterocycloalkyl, and heterocycloalkylalkyl and R¹ and R², together withthe nitrogen to which they attach, form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith between 0 and 3 R⁶ groups;

R³ is chosen from alkylamino, cycloalkylamino, arylamino,heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, andheterocycloalkylalkyl any of which may be optionally substituted withbetween 0 and 3 R⁶ groups;

R⁴, R^(4a), and R^(4b) are independently chosen from hydrogen, alkyl,alkenyl, alkynyl, and cycloalkyl;

R⁵ is chosen from aryl and heteroaryl, any of which may be optionallysubstituted with between 0 and 3 R⁶ groups;

each R⁶ is independently chosen from hydrogen, halogen, alkyl,alkylsulfonylaryl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy,haloaryl, alkoxyaryl, aryl, aryloxy, aralkyl, heterocycloalkyl,heteroaryl, alkylheteroaryl, heteroarylalkyl, cyano, alkoxy, alkoxyaryl,amino, alkylamino, dialkylamino, oxo, COR⁷, SO₂R⁷, NHSO₂R⁷, NHSO₂NHR⁷,NHCOR⁷, NHCONHR⁷, CONHR⁷, and CONR⁷R⁸; and

R⁷ and R⁸ are independently chosen from hydrogen, aryl, and lower alkyl;or R⁷ and R⁸ may be taken together to form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith lower alkyl.

In certain embodiments, the compound has Formula Ia, Ib, Ic, or Id:

or a salt, polymorph, or solvate thereof, wherein all groups are asdefined for Formula I.

In certain embodiments, the compound has Formula II or III:

or a salt, polymorph, or solvate thereof, wherein all groups are asdefined for Formula I.

In certain embodiments, the compound has Formula IIa, IIb, IIc, or IId:

or a salt, polymorph, or solvate thereof, wherein all groups are asdefined for Formula I. All cis and trans isomers are contemplated.

In certain embodiments, Z is NR^(4b).

In certain embodiments, R^(4b) is chosen from methyl and hydrogen.

In certain embodiments, R^(4b) is hydrogen.

In certain embodiments, the alkyl, whether by itself or as a named partof another non-cyclic substituent, is C₁-C₈ alkyl.

In certain embodiments, R³ is aryl, which may be optionally substitutedwith between 0 and 3 R⁶ groups.

In certain embodiments, R³ is chosen from aryl and heteroaryl, either ofwhich is substituted with an R⁶ group called R^(6a), chosen fromheteroaryl, cyano, and S(O)₂N(CH₃)₂. In certain embodiments, R³ remainsoptionally substituted with 1-2 additional R⁶ groups.

In certain embodiments, m is 0; Y is CH₂; and n is chosen from 0, 1, and2.

In certain embodiments, m is 0; Y is CH₂; and n is 2.

In certain embodiments, R³ is chosen from phenyl and heteroaryl, eitherof which is substituted with an R⁶ group called R^(6a), chosen fromheteroaryl, cyano, and S(O)₂N(CH₃)₂.

In certain embodiments, R^(6a) is chosen from cyano, S(O)₂N(CH₃)₂,

In certain embodiments, R^(6a) is heteroaryl.

In certain embodiments, R^(6a) is chosen from:

In certain embodiments, R^(6a) is

In certain embodiments, R⁴ is hydrogen.

In certain embodiments, nitrogen-containing heterocycloalkyl orheteroaryl ring formed by R¹ and R² together with the nitrogen to whichthey are attached contains 3 to eight atoms.

In certain embodiments, R¹ and R² are taken together to form anitrogen-containing heterocycloalkyl, which may be optionallysubstituted with between 0 and 3 R⁶ groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl is:

In certain embodiments, the nitrogen-containing heterocycloalkyl isoptionally substituted with between 0 and 3 R⁶ groups chosen from alkyland oxo.

In certain embodiments, R⁵ is phenyl, which may be optionallysubstituted with between 0 and 3 R⁶ groups.

In certain embodiments, R⁵ is:

wherein R^(6b) is chosen from halogen and hydroxy.

In certain embodiments, R^(6b) is chosen from fluoro, methoxy, andhydroxy.

In certain embodiments, R^(6b) is fluoro.

In certain embodiments, R^(4b) is chosen from methyl and hydrogen.

In certain embodiments, R^(4b) is hydrogen.

In certain embodiments, the compound has Formula IV:

or a salt, polymorph, or solvate thereof, wherein:

Y is chosen from a bond, NR^(4a), O, C(O)NH, NHC(O), S, SO₂, CHOH, andCH₂;

Z is chosen from a bond, NR^(4b), O, C(O)NH, NHC(O), S, SO₂, and CH₂;

m is chosen from 0, 1, 2, 3, 4, and 5;

n is chosen from 0, 1, 2, and 3;

R¹ and R² are each independently chosen from alkyl, aminoalkyl,alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl,cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl,heterocycloalkyl, and heterocycloalkylalkyl and R¹ and R², together withthe nitrogen to which they attach, form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith between 0 and 3 R⁶ groups;

R³ is chosen from alkylamino, cycloalkylamino, arylamino,heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, andheterocycloalkylalkyl any of which may be optionally substituted withbetween 0 and 3 R⁶ groups;

R^(4a) and R^(4b) are independently chosen from hydrogen, alkyl,alkenyl, alkynyl, and cycloalkyl; R⁵ is chosen from aryl and heteroaryl,any of which may be optionally substituted with between 0 and 3 R⁶groups;

R^(6a) is chosen from heteroaryl, cyano, and S(O)₂N(CH₃)₂;

each R⁶ is independently chosen from hydrogen, halogen, alkyl,alkylsulfonylaryl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy,haloaryl, alkoxyaryl, aryl, aryloxy, aralkyl, heterocycloalkyl,heteroaryl, alkylheteroaryl, heteroarylalkyl, cyano, alkoxy, alkoxyaryl,amino, alkylamino, dialkylamino, oxo, COR⁷, SO₂R⁷, NHSO₂R⁷, NHSO₂NHR⁷,NHCOR⁷, NHCONHR⁷, CONHR⁷, and CONR⁷R⁸; and

R⁷ and R⁸ are independently chosen from hydrogen, aryl, and lower alkyl;or R⁷ and R⁸ may be taken together to form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith lower alkyl.

In certain embodiments, Z is NR^(4b).

In certain embodiments, R^(4b) is chosen from methyl and hydrogen.

In certain embodiments, R^(4b) is hydrogen.

In certain embodiments, the alkyl, whether by itself or as a named partof another non-cyclic substituent, is C₁-C₈ alkyl.

In certain embodiments, m is 0; Y is CH₂; and n is chosen from 0, 1, and2.

In certain embodiments, n is 2.

In certain embodiments, R¹ and R² are each independently chosen fromalkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, and heteroaryl, andR¹ and R², together with the nitrogen to which they attach, form anitrogen-containing heterocycloalkyl or heteroaryl ring, which may beoptionally substituted with between 0 and 3 R⁶ groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl orheteroaryl ring formed by R¹ and R² together with the nitrogen to whichthey are attached contains 3 to eight atoms.

In certain embodiments, R¹ and R² are taken together to form anitrogen-containing heterocycloalkyl, which may be optionallysubstituted with between 0 and 3 R⁶ groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl isoptionally substituted with between 0 and 3 R⁶ groups chosen from alkyl,alkoxy, alkoxyalkyl, halogen, haloalkyl, haloalkoxy, hydroxy,hydroxyalkyl, alkylsulfonyl, alkylsulfonylalkyl, deuterium,trideuteromethyl, amino, —COOH, —CONH₂, —SO₂CH³, cyano,spiro-heterocycloalkyl, heteroaryl, and oxo.

In certain embodiments, the nitrogen-containing heterocycloalkyl isoptionally substituted with between 0 and 3 R⁶ groups chosen from alkyl,halogen, CONH₂, SO₂CH³, cyano, spiro-heterocycloalkyl, and oxo.

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl is:

In certain embodiments, R^(6a) is heteroaryl.

In certain embodiments, R^(6a) is chosen from cyano, S(O)₂N(CH₃)₂,

In certain embodiments, R^(6a) is chosen from

In certain embodiments, R⁵ is phenyl, which may be optionallysubstituted with between 0 and 3 R⁶ groups.

In certain embodiments, R⁵ is:

wherein R^(6b) is chosen from halogen, hydroxy, and methoxy.

In certain embodiments, R^(6b) is chosen from fluoro, methoxy, andhydroxy.

In certain embodiments, R^(6b) is fluoro.

In certain embodiments, the compound has Formula V:

or a salt, polymorph, or solvate thereof, wherein:

R¹ and R² are each independently chosen from alkyl, aminoalkyl,alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl,cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl,heterocycloalkyl, and heterocycloalkylalkyl and R¹ and R², together withthe nitrogen to which they attach, form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith between 0 and 3 R⁶ groups;

R^(4b) is chosen from hydrogen, alkyl, alkenyl, alkynyl, and cycloalkyl;

R^(6a) is chosen from heteroaryl, cyano, and S(O)₂N(CH₃)₂;

each R⁶ and each R^(6b) is independently chosen from hydrogen, halogen,alkyl, alkylsulfonylaryl, alkenyl, alkynyl, cycloalkyl, haloalkyl,haloalkoxy, haloaryl, alkoxyaryl, aryl, aryloxy, aralkyl,heterocycloalkyl, heteroaryl, alkylheteroaryl, heteroarylalkyl, cyano,alkoxy, alkoxyaryl, amino, alkylamino, dialkylamino, oxo, COR⁷, SO₂R⁷,NHSO₂R⁷, NHSO₂NHR⁷, NHCOR⁷, NHCONHR⁷, CONHR⁷, and CONR⁷R⁸; and

R⁷ and R⁸ are independently chosen from hydrogen, aryl, and lower alkyl;or R⁷ and R⁸ may be taken together to form a nitrogen-containingheterocycloalkyl or heteroaryl ring, which may be optionally substitutedwith lower alkyl.

In certain embodiments, R^(4b) is chosen from methyl and hydrogen.

In certain embodiments, R^(4b) is hydrogen.

In certain embodiments, R^(6a) is chosen from cyano, S(O)₂N(CH₃)₂,

In certain embodiments, R¹ and R² are each independently chosen fromalkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, and heteroaryl, andR¹ and R², together with the nitrogen to which they attach, form anitrogen-containing heterocycloalkyl or heteroaryl ring, which may beoptionally substituted with between 0 and 3 R⁶ groups

In certain embodiments, the nitrogen-containing heterocycloalkyl orheteroaryl ring formed by R¹ and R² together with the nitrogen to whichthey are attached contains 3 to eight atoms.

In certain embodiments, R¹ and R² are taken together to form anitrogen-containing heterocycloalkyl, which may be optionallysubstituted with between 0 and 3 R⁶ groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl ischosen from:

In certain embodiments, the nitrogen-containing heterocycloalkyl is:

In certain embodiments, R^(6b) is chosen from fluoro, methoxy, andhydroxy.

In certain embodiments, R^(6b) is fluoro.

Also provided are embodiments wherein any embodiment above may becombined with any one or more of these embodiments, provided thecombination is not mutually exclusive. As used herein, two embodimentsare “mutually exclusive” when one is defined to be something whichcannot overlap with the other. For example, an embodiment wherein Y isCH₂ is mutually exclusive with an embodiment wherein Y is NR^(4b).However, an embodiment wherein R¹ and R² are taken together to form anitrogen-containing heterocycloalkyl is not mutually exclusive with anembodiment wherein R⁵ is phenyl optionally substituted with fluorine.

In certain embodiments, the compound is chosen from the Examplesdisclosed herein, or a salt, polymorph, or solvate thereof. In certainembodiments, the compound is chosen from the Examples disclosed herein,or a salt, polymorph, or solvate thereof, wherein R^(6a) is chosen fromheteroaryl, cyano, and S(O)₂N(CH₃)₂. In certain embodiments, thecompound is chosen from the Examples disclosed herein, or a salt,polymorph, or solvate thereof, wherein R^(6a) is chosen from heteroaryland cyano. In certain embodiments, the compound is chosen from theExamples disclosed herein, or a salt, polymorph, or solvate thereof,wherein R^(6a) is heteroaryl.

TABLE 1 Compound Examples Ex # Structure Name 1

N-[3-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4- (1H-pyrazol-1-yl)benzamide 2

N-[3-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4- (pyrimidin-2-yl)benzamide 3

N-[3-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4- cyanobenzamide 4

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)biphenyl-4-carboxamide 5

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxobutan-2- yl)biphenyl-4-carboxamide 6

N-((S)-1-(azetidin-1-yl)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxobutan-2-yl)biphenyl-4-carboxamide 7

4′-fluoro-N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)biphenyl-4-carboxamide8

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)biphenyl-3-carboxamide 9

2-(biphenyl-4-yl)-N-((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4- (methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)acetamide 10

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)-2-(naphthalen-2-yl)acetamide 11

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)quinoline-3-carboxamide12

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)-4-(1H-pyrazol-1-yl)benzamide trifluoroacetic acid salt 13

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)-4-phenoxybenzamide 14

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)-1-naphthamide 15

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)-2-naphthamide 16

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)-4-(trifluoromethyl)benzamide 17

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)biphenyl-3-carboxamide18

N-((S)-4-((1R,2S)-2-(4- methoxyphenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)biphenyl-4-carboxamide19

N-((S)-4-((1R,2S)-2-(4- methoxyphenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan- 2-yl)biphenyl-4-carboxamide20

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)-4′-(methylsulfonyl)biphenyl- 4-carboxamide 21

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)-4-(oxazol-2-yl)benzamide22

N-((2S)-4-(2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)-(2′,3′,4′,5′,6′-d₅)-biphenyl-4- carboxamide 23

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]aminol-1-oxo-1-(1H-1,2,3-triazin-1-yl)pentan-2-yl]-4- (pyrimidin-2-yl)benzamide 24

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(azetidin-4-yl)-1-oxopentan-2-yl]-4-(1H- pyrazol-1-yl)benzamidetrifluoroacetic acid sail 25

N-[(2S)-1-[3-(dimethylamino)pyrrolidin-1-yl]-5-[[(1R,2S)-2-(4-fluorophenyl)- cyclopropyl]amino)-1-oxopentan-2-yl]-4- (1H-1,2,3-triazol-1-yl)benzamide 26

N-[(2S)-1-[4-(dimethylamino)piperidin-1-yl]-5-[[(1R,2S)-2-(4-fluorophenyl)- cyclopropyl]amino]-1-oxopentan-2-yl]-4- (1H-1,2,3-triazol-1-yl)benzamide 27

N-[(2S)-5-[((1R,2S)-2-(4- fluorophenyl)cyclopropyl] amino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4- (1,2,4-triazol-4-yl)benzamide28

N-[(2S)-5-[[1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-[4-(2-hydroxyethyl)piperazin-1-yl]-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 29

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(azetidin-1-yl)-1-oxopentan-2-yl]-4- cyanobenzamide 30

1-((S)-2-(4-cyanobenzamido)-5-((1R,2S)-2- (4-fluorophenyl)cyclopropylamino)pentanoyl)-4-fluoropiperidine-4-carboxamide 31

Ethyl 2-[4-[(2S)-2-[(4- cyanophenyl)formamido]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl] amino]pentanoyl]piperazin-1-yl]acetate 32

4-cyano-N-[(2S)-5-[[1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(pyrimidin-2-yl)piperazin-1-yl]pentan-2- yl]benzamide 33

4-fluoro-1-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-2-(4-(pyrimidin-2- yl)benzamido)pentanoyl)piperidine-4- carboxamide 34

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-[2-oxa-6-azaspiro[3.3]heptan-6-yl]-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 35

N-[(2S)-1-(3-cyanoazetidin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]-4-(1H-pyrazol-1-yl)benzamide 36

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-pyrazol-1-yl)benzamide 37

1-((S)-2-(4-(1H-pyrazol-1-yl)benzamido)-5- ((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)pentanoyl)-4-fluoropiperidine-4-carboxamide 38

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-[2-oxa-6-azaspiro[3.3]heptan-6-yl]-1-oxopentan-2-yl]-4-(1H-pyrazol-1-yl)benzamide 39 This example is intentionally leftblank. 40 This example is intentionally left blank. 41

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-oxo-1-[(3R,5S)-3,4,5-trimethylpiperazin-1-yl]pentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 42

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl] amino]-1-(2-cyanopyrrolidine-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 43

N-[(2S)-1-(4,4-difluoropiperidin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 44

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-cyanopiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 45

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methanesulfonylpiperidin-1-yl)-1- oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 46

N-[(2S)-1-(4-Ethylpiperazin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 47

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(propan-2-yl)piperazin-1-yl]pentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 48

N-[(2S)-5-[[(2S)-1-(4-fluorophenyl)-1{circumflex over ( )}[3]-oxiran-2-yl]amino]-1-(4- methanesulfonylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 49

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl] amino]-1-[4-(2-hydroxyethyl) 1H-1,2,3-triazol-1-yl]-1- oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 50

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino)-1-(4-(2-methoxyethyl)-piperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 51

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl] amino]-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxopentan-2-yl]-4-(1,2,3-triazol--yl)benzamide 52

N-[(2S)-5-[[(1R-2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-(3-oxopiperazin-1-yl)pentan-2-yl]-4-(1H- 1,2,3-triazol-1-yl)benzamide 53

N-[(2S)-1-[(2R,6S)-2,6-dimethylmorpholin- 4-yl]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 54

N-[(2S)-1-[3-Azabicyclo[3.2.1] octan-3-yl]- 5-[[(2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 55

N-[(2S)-5-[[(1R,2S)-2-(4- Fluoropheny)cyclopropyl] amino]-1-(8-methyl-3,8-diazabicyclo[3.2.1]octane-l-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 56

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(8-oxa-3-azabicyclo[3.2.1]octane-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 57

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(2-methyl-2,8- diazaspiro[4.5]decane-8-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 58

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(8-methyl-2,8- diazaspiro[4.5]decane-2-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 59

N-((2S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 60

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]pentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 61

1-((S)-2-(4-(1H-1,2,3-triazol-1- yl)benzamido)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)pentanoyl)-4-fluoropiperidine-4-carboxamide 62

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-(3-oxopiperazin-1-yl)pentan-2-yl]-4-(4H- 1,2,4-triazol-4-yl)benzamide 63

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxopentan-2-yl]-4-(4H-1,2,4-triazol-4-yl)benzamide 64

N-[(2S)-1-(4-ethylpiperazin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]-4-fluorobenzamide;trifluoroacetic acid 65

4-cyano-N-[(2S)-1-(3,3-dimethylazetidin-1- yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]benzamide 66

4-cyano-N-((S)-1-(3-cyanoazetidin-1-yl)-5- ((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxopentan-2-yl)benzamide 67

4-cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]aminol-1-[3-(2-hydroxypropan-2-yl)azetidin-1-yl]-1- oxopentan-2-yl]benzamide 68

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-hydroxy-4-methylpiperidin-1-yl)-1- oxopentan-2-yl]benzamide 69

4-Cyano-N-[(2S)-5-[[(2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methoxy-4-methylpiperidin-1-yl)-1- oxopentan-2-yl]benzamide 70

4-Cyano-N-[(2S)-5-[[(2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(1H-pyrazol-1-yl)piperidin-1-yl]pentan- 2-yl]benzamide 71

4-cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(2,2,2-trifluoroethyl)piperazin-1- yl]pentan-2-yl]benzamide 72

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(1H-1,2,3-triazol-1-yl)piperidin-1- yl]pentan-2-yl]benzamide 73

4-Cyano-N-[(2S)-5-[[(2S)-2-(4- fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-oxo-1-[4-(1H-1,2,4-triazol-4-yl)piperidin-1-yl]pentan-2-yl]benzamide 74

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(1H-1,2,3,4-tetrazol-5-yl)piperidin-1- yl]pentan-2-yl]benzamide 75

4-Cyano-N-[(2S)-1-(4-ethylpiperazin-1-yl)- 5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]benzamide 76

4-Cyano-N-[(2S)-5-[[(2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(propan-2-yl)piperazin-1-yl]pentan-2- yl]benzamide 77

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-(2-hydroxyethyl)piperazin-1-yl]-1-oxopentan-2-yl]benzamide 78

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-[4-(2-methoxyethyl)piperazin-1-yl)-1-oxopentan- 2-yl]benzamide 79

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-(2- methanesulfonylethyl)piperazin-1-yl]-1-oxopentan-2-yl]benzamide 80

4-Cyano-N-[(2S)-1-(4-ethyl-3- oxopiperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]benzamide 81

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(2,4,5,6-tetrahydro-2-methyl-pyrrolo[3,4-c]pyrazol-1-yl)-1-oxopentan-2-yl]-4- cyanobenzamide 82

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(2,4,5,6-tetrahydro-2- (methanesulfonyl)pyrrolo[3,4-c]pyrazol-1-yl)-1-oxopentan-2-yl]-4-cyanobenzamide 83

4-cyano-N-((2S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pentan-2- yl)benzamide 84

4-cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-[2-oxa-8-azaspiro[4.5]decan-8-yl]-1-oxopentan-2- yl]benzamide 85

N-[(2S)-1-(4-Ethylpiperazin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 86

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-oxo-1-(piperazin-1-yl)pentan-2-yl)-4-(pyrimidin-2- yl)benzamide 87 Thisexample is intentionally left blank. 88 This example is intentionallyleft blank. 89

N-((S)-1-((3S,5R)-3,5-dimethylpiperazin-1- yl)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 90

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(3,8-diazabicyclo[3.2.1]octane-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 91

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-carboxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 92

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl] amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2- yl]benzamide 93

N-[(2S)-5-[[(2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]- 4-(pyrimidin-2-yl)benzamide 94

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2- yl]benzamide 95

4-Methanesulfonyl-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methyl-3-oxopiperazin-1-yl]-1-oxopentan- 2-yl]benzamide 96

4-(Dimethylsulfamoyl)-N-[(2S)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2- yl]benzamide 97

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl] amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4- (1-pyrrolyl)benzamide 98

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(2H-1,2,3-triazol-2-yl)benzamide 99

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(2-methyl-2H-1,2,3-triazol-4-yl)benzamide 100

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,4-triazol-1-yl)benzamide 101

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1-methyl-1H-1,2,4-triazol-3-yl)benzamide 102

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]aminol-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3,4-tetrazol-5-yl)benzamide 103

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4- (pyrimidin-2-yl)benzamide 104

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-2-phenylpyrimidine-5-carboxamide 105

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-6-methoxynaphthalene-2-carboxamide 106

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(propan-2-yl)piperaxin-1-yl]pentan-2-yl]- 4-(pyrimidin-2-yl)benzamide107

N-[(2S)-1-[4-(2-Cyanoacetyl)piperazin-1- yl]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 108

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino)-1-(1,4-diazabicyclo[3.2.2]nonane-1-yl)-1- oxopentan-2-yl]-4-cyanobenzamide 109

4-[[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2- yl]carbamoyl]benzoic acid 110

N-[(2R)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 111

N-[(2S)-5-[[(2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 112

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4- (1H-imidazol-1-yl)benzamide113

4-Fluoro-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-[4-(propan-2-yl)piperazin-1-yl]pentan-2- yl]benzamide 114

6-cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino)-1-oxo-1-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]pentan-2-yl]pyridine-3-carboxamide 115

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl] amino]-1-(thiomorpholine-4,4-dioxide-1-yl)-1- oxopentan-2-yl]-4-(pyrrolidin-1-yl)benzamide 116

N-[(2S)-1-(3-cyanoazetidin-1-yl)-5- [[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]-4-(1H-imidazol-1-yl)benzamide 117

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(3-(dimethylamino)-azetidin-1-yl-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)- benzamide 118

N-[(2S)-1-[4-(2-Cyanoacetyl)piperazin-1- yl]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1- oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 119

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3,4-tetrazol-1-yl)benzamide 120

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3,4-tetrazol-1-yl)benzamide 121

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1- (3-oxopiperazin-1-yl)pentan-2-yl]benzamide 122

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxopentan- 2-yl]benzamide 123

N-[(2S)-1-[1,4-Diazabicyclo[3.2.2]nonan-4- yl]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl] amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 124

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino)-1-(1,4-diazabicyclo[3.2.2]nonane-1-yl)-1- oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide 125

4-cyano-N-((S)-1-(3,3-difluoroazetidin-1- yl)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxopentan-2-yl)benzamide 126

2-[4-[(2S)-2-[(4-Cyanophenyl)formamido]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]-amino]pentanoyl]piperazin-1-yl]acetic acid 127

benzyl N-[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]-N-[(4R)-5-(4-methylpiperazin-1-yl)-5-oxo-4-[[4-(1H- 1,2,3-triazol-l-yl)phenyl)formamido]pentyl]carbamate 128

N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methanesulfonylpiperazin-1-yl)-1- oxopentan-2-yl]-4-phenylbenzamide 129

N-[(2S)-5-[[(1R,2S)-2-(4- methoxyphenyl)cyclopropyl] amino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 130

2-Fluoro-N-[(2S)-5-[[(1R,2S)-2-(4- fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 131

N-[(2S)-1-(4-cyano-4-fluoropiperidin-1-yl)- 5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1- yl)benzamide 132

N-((S)-5-((1S,2R)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 133

N-((R)-5-((1S,2R)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 134

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl] amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4- (pyrazin-2-yl)benzamide 135

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4- (pyrimidin-5-yl)benzamide 136

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4- (pyridazin-3-yl)benzamide 137

N-[(2S)-5-[[(2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-[4-(d₃)-methylpiperazin-1-yl]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide 138

N-[(2S)-5-[[(1R,2S)-2-(4- Fluorophenyl)cyclopropyl]amino]-1-(4-methyl-(2,2,3,3,5,5,6,6-d₈)-piperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)- benzamide 139

1-((R)-2-(4-(1H-pyraxol-1-yl)benzamido)-5- ((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)pentanoyl)-4-fluoropiperidine-4-carboxamide 140

N-((R)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4- (1H-imidazol-1-yl)benzamide141

N-((S)-6-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxohexan-2-yl)biphenyl-4-carboxamide142

N-((S)-6-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxohexan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 143

4-cyano-N-((S)-6-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxohexan-2- yl)benzamide 144

N-((S)-6-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxohexan-2-yl)-4- (pyrimidin-2-yl)benzamide 145

4-fluoro-N-((R)-3-(2-((1R,2S)-2-(4- fluorophenyl)-1-methylcyclopropylamino)ethylsulfonyl)-1-morpholino-1-oxopropan-2-yl)benzamide 146

3,4-dichloro-N-((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4- (methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)benzamide 147

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)-2-(4′-methoxybiphenyl-4- yl)acetamide 148

4-(2,5-(dimethyl-1H-pyrrol-1-yl)-N-((S)-4- ((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)benzamide 149

3,4-dichloro-N-((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4- (methylsulfonyl(piperdzin-1-yl)-1-oxobutan-2-yl)benzamide 150

N-((S)-4-((1S,2R)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)biphenyl-4-carboxamide 151

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2- yl)biphenyl-4-carboxamide152

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)-4′-(methylsulfonyl)biphenyl-4- carboxamide 153

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2- yl)biphenyl-4-carboxamide 154

N-((S)-4-((1S,2R)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxobutan-2- yl)biphenyl-4-carboxamide 155

4-fluoro-N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxobutan-2- yl)benzamide 156

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(1H-pyrazol-1-yl)benzamide 157

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4- (1H-pyrazol-1-yl)benzamide158

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 159

N-((S)-4-((1R,2S)-2-(4- fluorophenl)cyclopropylamino)-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxobutan-2- yl)biphenyl-4-carboxamide 160

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(3-oxopiperazin-1-yl)butan-2-yl)biphenyl-4- carboxamide 161

4-fluoro-N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2- yl)benzamide 162

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(pyridin-2-yl)benzamide 163

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(4H-1,2,4-triazol-4-yl)benzamide 164

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide 165

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2- yl)picolinamide 166

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-1-phenyl-1H-pyrazole-4-carboxamide 167

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(1H-imidazol-1-yl)benzamide 168

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(pyrimidin-2-yl)benzmide 169

4-fluoro-N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(3-oxopiperazin-1-yl)pentan-2- yl)benzamide 170

4-fluoro-N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methyl-3-oxopiperazin-1-yl)-1-oxopentan- 2-yl)benzamide 171

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4′-(methylsulfonyl)biphenyl-4- carboxamide 172

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2- yl)-4-(oxazol-2-yl)benzamide173

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(pyrazin-2-yl)benzamide 174

4-cyano-N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2- yl)benzamide 175

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(3-oxopiperazin-1-yl)pentan-2-yl)-4-(1H- pyrazol-1-yl)benzamide 176

N-((S)-1-(azetidin-1-yl)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxopentan-2-yl)-4-(1H-pyrazol-1-yl)benzamide 177

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-morpholino-1-oxopentan-2-yl)-4-(1H- pyrazol-1-yl)benzamide 178

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxopentan-2-yl)-4-(1H-pyrazol-1-yl)benzamide 179

4-fluoro-N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-morpholino-1-oxopentan-2-yl)benzamide 180

N-((S)-5-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(1H-pyrrol-1-yl)benzamide 181

N-((S)-5-((1R,2S)-2-(4- fluorophenyl(cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4- (pyrimidin-2-yl)benzamide 182

N-((S)-3-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl)-4- (1H-pyrazol-1-yl)benzamidedihydrochloride sail 183

N-((S)-4-((1S,2R)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)-6-methoxy-2-naphthamide 184

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxobutan-2-yl)biphenyl-4-carboxamide185

N-((S)-6-((1S,2R)-2-(4-fluorophenyl)-1- methylcyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1- oxohexan-2-yl)benzamide 186

N-((S)-4-((1R,2S)-2-(4- fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)-4-(phenylsulfonyl)benzamide 187

4′-fluoro-N-((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2- yl)biphenyl-4-carboxamide 188

N-((S)-1-(azetidin-1-yl)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1- oxopentan-2-yl)-4-fluorobenzamide

Also provided herein is salt of a compound as disclosed herein, or apolymorph or solvate thereof.

In certain embodiments, the salt has Formula VI:

or a polymorph or solvate thereof, wherein:

X is chosen from tosylate, sulfate, tartrate, oxalate, besylate,fumarate, citric, esylate, and malate; and

q is an integer chosen from 1 and 2.

In certain embodiments, X is tosylate.

In certain embodiments, q is 2.

Provided herein isN—((S)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamidebis-tosylate, or a polymorph thereof. Also provided herein isN—((S)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamidebis-tosylate Form 2.

Also provided herein is a tosylate salt of a compound as disclosedherein, or a polymorph or solvate thereof. Also provided herein is abis-tosylate salt of a compound as disclosed herein, or a polymorph orsolvate thereof.

Also provided herein is a compound, or a salt, polymorph, or solvatethereof, as disclosed herein is provided for use as a medicament. Alsoprovided herein is a compound as disclosed herein, or a salt, polymorph,or solvate thereof, for use in the manufacture of a medicament for theprevention or treatment of a KDM1A-mediated disease.

Also provided herein is a compound as disclosed herein is for use in themanufacture of a medicament for the prevention or treatment of a diseaseor condition chosen from sickle cell disease, thalassemia major, andother beta-hemoglobinopathies.

Also provided herein is a pharmaceutical composition is provided whichcomprises a compound as disclosed herein, together with apharmaceutically acceptable carrier.

In certain embodiments, the pharmaceutical composition is formulated fororal administration.

In certain embodiments, the pharmaceutical composition additionallycomprises another therapeutic agent.

Also provided herein is a method of inhibiting KDM1A is provided,comprising contacting KDM1A with a compound as disclosed herein.

Also provided herein is a method of treatment of a KDM1A-mediateddisease comprising the administration of a therapeutically effectiveamount of a compound as disclosed herein, or a salt, polymorph, orsolvate thereof, to a patient in need thereof.

In certain embodiments, the disease is cancer.

In certain embodiments, the cancer is chosen from Ewing's sarcoma,multiple myeloma, T-cell leukemia, Wilm's tumor, small-cell lung cancer,bladder cancer, prostate cancer, breast cancer, head/neck cancer, coloncancer, and ovarian cancer.

In certain embodiments, the disease is a myeloid disease.

In certain embodiments, the myeloid disease is chosen frommyelofibrosis, polycythemia vera, essential thrombocythemia,myelodysplastic syndrome (MDS), acute myelogenous leukemia (AML), andchronic myelogenous leukemia (CML).

In certain embodiments, the disease is an inflammatory disease.

In certain embodiments, the inflammatory disease is chosen frominflammatory bowel disease, rheumatoid arthritis, or systemic lupuserythematosus.

Also provided herein is a method of treatment of a globin-mediateddisease comprising the administration of a therapeutically effectiveamount of a compound as disclosed herein, or a salt, polymorph, orsolvate thereof, to a patient in need thereof.

Also provided herein is a method for achieving an effect in a patient isprovided; comprising the administration of a therapeutically effectiveamount of a compound as disclosed herein; wherein the effect is chosenfrom an elevation of red blood cell count, an elevation of the red bloodcell count of red cells containing fetal hemoglobin, an elevation in thetotal concentration of fetal hemoglobin in red cells, an elevation inthe total concentration of fetal hemoglobin in reticulocytes, anincrease in the transcription of the gamma globin gene in bonemarrow-derived red cell precursors, e.g., pro-erythroblasts, a reductionin the number of sickle cell crises a patient experiences over a unitperiod of time, a halt to or prevention of tissue damage e.g. in theheart, spleen, brain or kidney caused by sickling cells, a reduction inthe proportion of red cells that undergo sickling under physiologicalconditions of relative hypoxia as measured using patient blood in an invitro assay, an increase in the amount of histone 3 lysine methylationat lysine position 4 (H3K4me1 and H3K4me2), and/or a decrease in theamount of histone 3 methylation at lysine position 9 (H3K9me1 orH3K4me2) near or at the gamma globin promoter as assayed by ChIP usingcells derived from a treated patient.

Also provided herein is a method of inhibiting at least one KDM1Afunction is provided; comprising the step of contacting KDM1A with acompound as disclosed herein; wherein the inhibition is measured byphenotype of red cells or their precursors either cultured or in vivo inhumans or mouse or transgenic mice containing the human beta globinlocus or portions thereof, the ability of cancer cells to proliferate,the expression of specific genes known to be regulated by KDM1A activitysuch as gamma globin, a change in the histone methylation states, achange in the methylation state of proteins known to be demethylated byKDM1A such as G9a or SUV39H1, expression of KDM1A-regulated genes, orbinding of KDM1A with a natural binding partner such as CoREST, DNMT1 orHDACs.

Inhibition of KDM1A (LSD1) activity alone may be sufficient therapy forthe treatment of some diseases; for other such as cancer, combinationtherapies are often additive or synergistic in their therapeutic effectsand may even be necessary to achieve the full clinical benefit desired.There is specific scientific evidence to rationalize the combination ofan inhibitor of KDM1A with all-trans retinoic acid (ATRA), arsenictrioxide, inhibitors of DNA methyltransferases such as 5′-azacytidine or5′-aza 2′-deoxycytidine, inhibitors of NFκB signaling such as sulindacor conventional anti-neoplastic agents such as anthracyclines ornucleoside analogues such as cytosine arabinoside. Likewise, agents thatinduce leukemia stem cells into the cell cycle (G-CSF, GM-CSF, stem cellfactor, thrombopoietin (TPO)) or agents that negate the contributoryrole cytokines (TPO, CCL3(MIP-1)) play in remodeling the niche of cancerstem cells may be useful as part of a combination including an LSD1inhibitor.

Abbreviations and Definitions

To facilitate understanding of the disclosure, a number of terms andabbreviations as used herein are defined below as follows:

When introducing elements of the present disclosure or the preferredembodiment(s) thereof, the articles “a”, “an”, “the” and “said” areintended to mean that there are one or more of the elements. The terms“comprising”, “including” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

The term “and/or” when used in a list of two or more items, means thatany one of the listed items can be employed by itself or in combinationwith any one or more of the listed items. For example, the expression “Aand/or B” is intended to mean either or both of A and B, i.e. A alone, Balone or A and B in combination. The expression “A, B and/or C” isintended to mean A alone, B alone, C alone, A and B in combination, Aand C in combination, B and C in combination or A, B, and C incombination.

The term “about,” as used herein when referring to a measurable valuesuch as an amount of a compound, dose, time, temperature, and the like,is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1%from the specified amount.

A “therapeutically effective amount” of a drug is an amount of drug orits pharmaceutically acceptable salt that eliminates, alleviates, orprovides relief of the symptoms of the disease for which it isadministered.

A “subject in need thereof” is a human or non-human animal that exhibitsone or more symptoms or indicia of a disease.

When ranges of values are disclosed, and the notation “from n₁ . . . ton₂” or “between n₁ . . . and n₂” is used, where n₁ and n₂ are thenumbers, then unless otherwise specified, this notation is intended toinclude the numbers themselves and the range between them. This rangemay be integral or continuous between and including the end values. Byway of example, the range “from 2 to 6 carbons” is intended to includetwo, three, four, five, and six carbons, since carbons come in integerunits. Compare, by way of example, the range “from 1 to 3 μM(micromolar),” which is intended to include 1 μM, 3 μM, and everythingin between to any number of significant figures (e.g., 1.255 μM, 2.1 μM,2.9999 μM, etc.). When n is set at 0 in the context of “0 carbon atoms”,it is intended to indicate a bond or null.

The term “alkylsulfonyl” as used herein, means an alkyl group, asdefined herein, appended to the parent molecular moiety through asulfonyl group, as defined herein. Representative examples ofalkylsulfonyl include, but are not limited to, methylsulfonyl andethylsulfonyl.

The term “alkylsulfonylalkyl” as used herein, means an alkylsulfonylgroup, as defined herein, appended to the parent molecular moietythrough an alkyl group, as defined herein. Representative examples ofalkylsulfonylalkyl include, but are not limited to, methylsulfonylmethyland ethylsulfonylmethyl.

The term “acyl,” as used herein, alone or in combination, refers to acarbonyl attached to an alkenyl, alkyl, aryl, cycloalkyl, heteroaryl,heterocycle, or any other moiety where the atom attached to the carbonylis carbon. An “acetyl” group refers to a —C(O)CH₃ group. An“alkylcarbonyl” or “alkanoyl” group refers to an alkyl group attached tothe parent molecular moiety through a carbonyl group. Examples of suchgroups include methylcarbonyl and ethylcarbonyl. Examples of acyl groupsinclude formyl, alkanoyl and aroyl.

The term “alkenyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon group having one or moredouble bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkenyl will comprise from 2 to 6 carbon atoms. Theterm “alkenylene” refers to a carbon-carbon double bond system attachedat two or more positions such as ethenylene [(—CH═CH—), (—C::C—)].Examples of suitable alkenyl groups include ethenyl, propenyl,2-methylpropenyl, 1,4-butadienyl and the like. Unless otherwisespecified, the term “alkenyl” may include “alkenylene” groups.

The term “alkoxy,” as used herein, alone or in combination, refers to analkyl ether group, wherein the term alkyl is as defined below. Examplesof suitable alkyl ether groups include methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, and the like.

The term “alkyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain alkyl group containing from 1 to 20carbon atoms. In certain embodiments, said alkyl will comprise from 1 to10 carbon atoms. In further embodiments, said alkyl will comprise from 1to 6 carbon atoms. Alkyl groups may be optionally substituted as definedherein. Examples of alkyl groups include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,hexyl, octyl, noyl and the like. The term “alkylene,” as used herein,alone or in combination, refers to a saturated aliphatic group derivedfrom a straight or branched chain saturated hydrocarbon attached at twoor more positions, such as methylene (—CH₂—). Unless otherwisespecified, the term “alkyl” may include “alkylene” groups.

The term “alkylamino,” as used herein, alone or in combination, refersto an alkyl group attached to the parent molecular moiety through anamino group. Suitable alkylamino groups may be mono- or dialkylated,forming groups such as, for example, N-methylamino, N-ethylamino,N,N-dimethylamino, N,N-ethylmethylamino and the like.

The term “alkylidene,” as used herein, alone or in combination, refersto an alkenyl group in which one carbon atom of the carbon-carbon doublebond belongs to the moiety to which the alkenyl group is attached.

The term “alkylthio,” as used herein, alone or in combination, refers toan alkyl thioether (R—S—) group wherein the term alkyl is as definedabove and wherein the sulfur may be singly or doubly oxidized. Examplesof suitable alkyl thioether groups include methylthio, ethylthio,n-propylthio, isopropylthio, n-butylthio, iso-butylthio, sec-butylthio,tert-butylthio, methanesulfonyl, ethanesulfinyl, and the like.

The term “alkynyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon group having one or moretriple bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkynyl comprises from 2 to 6 carbon atoms. In furtherembodiments, said alkynyl comprises from 2 to 4 carbon atoms. The term“alkynylene” refers to a carbon-carbon triple bond attached at twopositions such as ethynylene (—C≡C—). Examples of alkynyl groups includeethynyl, propynyl, hydroxypropynyl, butyn-1-yl, butyn-2-yl, pentyn-1-yl,3-methylbutyn-1-yl, hexyn-2-yl, and the like. Unless otherwisespecified, the term “alkynyl” may include “alkynylene” groups.

The terms “amido” and “carbamoyl,” as used herein, alone or incombination, refer to an amino group as described below attached to theparent molecular moiety through a carbonyl group, or vice versa. Theterm “C-amido” as used herein, alone or in combination, refers to a—C(═O)—NR₂ group with R as defined herein. The term “N-amido” as usedherein, alone or in combination, refers to a RC(═O)NH— group, with R asdefined herein. The term “acylamino” as used herein, alone or incombination, embraces an acyl group attached to the parent moietythrough an amino group. An example of an “acylamino” group isacetylamino (CH₃C(O)NH—).

The term “amino,” as used herein, alone or in combination, refers to—NRR′, wherein R and R′ are independently chosen from hydrogen, alkyl,hydroxyalkyl, acyl, heteroalkyl, aryl, cycloalkyl, heteroaryl, andheterocycloalkyl, any of which may themselves be optionally substituted.Additionally, R and R′ may combine to form heterocycloalkyl, either ofwhich may be optionally substituted.

The term “amino acid”, as used herein, alone or in combination, refersto a —NHCHRC(O)O— group, which may be attached to the parent molecularmoiety to give either an N-terminus or C-terminus amino acid, wherein Ris independently chosen from hydrogen, alkyl, aryl, heteroaryl,heterocycloalkyl, aminoalkyl, amido, amidoalkyl, carboxyl,carboxylalkyl, guanidinealkyl, hydroxyl, thiol, and thioalkyl, any ofwhich themselves may be optionally substituted. The term C-terminus, asused herein, alone or in combination, refers to the parent molecularmoiety being bound to the amino acid at the amino group, to give anamide as described herein, with the carboxyl group unbound, resulting ina terminal carboxyl group, or the corresponding carboxylate anion. Theterm N-terminus, as used herein, alone or in combination, refers to theparent molecular moiety being bound to the amino acid at the carboxylgroup, to give an ester as described herein, with the amino groupunbound resulting in a terminal secondary amine, or the correspondingammonium cation. In other words, C-terminus refers to —NHCHRC(O)OH or to—NHCHRC(O)O— and N-terminus refers to H₂NCHRC(O)O— or to H₃N⁺CHRC(O)O—.

The term “aminoalkyl,” as used herein, alone or in combination, refersto an amino group as defined herein linked through an alkyl group to theparent moiety.

The term “aryl”, as used herein, alone or in combination, means acarbocyclic aromatic system containing one, two or three rings whereinsuch polycyclic ring systems are fused together. The term “aryl”embraces aromatic groups such as phenyl, naphthyl, anthracenyl, andphenanthryl.

The term “arylalkenyl” or “aralkenyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkenyl group.

The term “arylalkoxy” or “aralkoxy,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkoxy group.

The term “arylalkyl” or “aralkyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkyl group.

The term “arylalkynyl” or “aralkynyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkynyl group.

The term “arylalkanoyl” or “aralkanoyl” or “aroyl,” as used herein,alone or in combination, refers to an acyl group derived from anaryl-substituted alkanecarboxylic acid such as benzoyl, naphthoyl,phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4-phenylbutyryl,(2-naphthyl)acetyl, 4-chlorohydrocinnamoyl, and the like.

The term aryloxy as used herein, alone or in combination, refers to anaryl group attached to the parent molecular moiety through an oxy.

The terms “benzo” and “benz,” as used herein, alone or in combination,refer to the divalent group C₆H₄=derived from benzene. Examples includebenzothiophene and benzimidazole.

The term “biphenyl” as used herein refers to two phenyl groups connectedat one carbon site on each ring.

The term “carbamate,” as used herein, alone or in combination, refers toan ester of carbamic acid (—NHCOO—) which may be attached to the parentmolecular moiety from either the nitrogen or acid end, and which may beoptionally substituted as defined herein.

The term “O-carbamyl” as used herein, alone or in combination, refers toa —OC(O)NRR′ group, with R and R′ as defined herein.

The term “N-carbamyl” as used herein, alone or in combination, refers toa ROC(O)NR′— group, with R and R′ as defined herein.

The term “carbonyl,” as used herein, when alone includes formyl [—C(O)H]and in combination is a —C(O)— group.

The term “carboxyl” or “carboxy,” as used herein, refers to —C(O)OH orthe corresponding “carboxylate” anion, such as is in a carboxylic acidsalt. An “O-carboxy” group refers to a RC(O)O— group, where R is asdefined herein. A “C-carboxy” group refers to a —C(O)OR groups where Ris as defined herein.

The term “cyano,” as used herein, alone or in combination, refers to—CN.

The term “cycloalkyl,” or, alternatively, “carbocycle,” as used herein,alone or in combination, refers to a saturated or partially saturatedmonocyclic, bicyclic or tricyclic alkyl group wherein each cyclic moietycontains from 3 to 12 carbon atom ring members and which may optionallybe a benzo fused ring system which is optionally substituted as definedherein. In certain embodiments, said cycloalkyl will comprise from 5 to7 carbon atoms. Examples of such cycloalkyl groups include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronaphthyl,indanyl, octahydronaphthyl, 2,3-dihydro-1H-indenyl, adamantyl and thelike. “Bicyclic” and “tricyclic” as used herein are intended to includeboth fused ring systems, such as decahydronaphthalene,octahydronaphthalene as well as the multicyclic (multicentered)saturated or partially unsaturated type. The latter type of isomer isexemplified in general by, bicyclo[1,1,1]pentane, camphor, adamantane,and bicyclo[3,2,1]octane.

The term “ester,” as used herein, alone or in combination, refers to acarboxy group bridging two moieties linked at carbon atoms.

The term “ether,” as used herein, alone or in combination, refers to anoxy group bridging two moieties linked at carbon atoms.

The term “guanidine”, as used herein, alone or in combination, refers to—NHC(═NH)NH₂, or the corresponding guanidinium cation.

The term “halo,” or “halogen,” as used herein, alone or in combination,refers to fluorine, chlorine, bromine, or iodine.

The term “haloalkoxy,” as used herein, alone or in combination, refersto a haloalkyl group attached to the parent molecular moiety through anoxygen atom.

The term “haloalkyl,” as used herein, alone or in combination, refers toan alkyl group having the meaning as defined above wherein one or morehydrogen atoms are replaced with a halogen. Specifically embraced aremonohaloalkyl, dihaloalkyl and polyhaloalkyl groups. A monohaloalkylgroup, for one example, may have an iodo, bromo, chloro or fluoro atomwithin the group. Dihalo and polyhaloalkyl groups may have two or moreof the same halo atoms or a combination of different halo groups.Examples of haloalkyl groups include fluoromethyl, difluoromethyl,trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl,pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl anddichloropropyl. “Haloalkylene” refers to a haloalkyl group attached attwo or more positions. Examples include fluoromethylene (—CFH—),difluoromethylene (—CF₂—), chloromethylene (—CHCl—) and the like.

The term “heteroalkyl,” as used herein, alone or in combination, refersto a stable straight or branched chain, or cyclic hydrocarbon group, orcombinations thereof, fully saturated or containing from 1 to 3 degreesof unsaturation, consisting of the stated number of carbon atoms andfrom one to three heteroatoms chosen from O, N, and S, and wherein thenitrogen and sulfur atoms may optionally be oxidized and the nitrogenheteroatom may optionally be quaternized. The heteroatom(s) O, N and Smay be placed at any interior position of the heteroalkyl group. Up totwo heteroatoms may be consecutive, such as, for example, —CH₂—NH—OCH₃.

The term “heteroaryl,” as used herein, alone or in combination, refersto a 3 to 7 membered unsaturated heteromonocyclic ring, or a fusedmonocyclic, bicyclic, or tricyclic ring system in which at least one ofthe fused rings is aromatic, which contains at least one atom chosenfrom O, S, and N. In certain embodiments, said heteroaryl will comprisefrom 5 to 7 carbon atoms. The term also embraces fused polycyclic groupswherein heterocyclic rings are fused with aryl rings, wherein heteroarylrings are fused with other heteroaryl rings, wherein heteroaryl ringsare fused with heterocycloalkyl rings, or wherein heteroaryl rings arefused with cycloalkyl rings. Examples of heteroaryl groups includepyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, triazolyl, pyranyl, furanyl, thienyl, oxazolyl,isoxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, isothiazolyl, indolyl,isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl,quinoxalinyl, quinazolinyl, indazolyl, benzotriazolyl, benzodioxolyl,benzopyranyl, benzoxazolyl, benzoxadiazolyl, benzothiazolyl,benzothiadiazolyl, benzofuranyl, benzothienyl, chromonyl, coumarinyl,benzopyranyl, tetrahydroquinolinyl, tetrazolopyridazinyl,tetrahydroisoquinolinyl, thienopyridinyl, furopyridinyl,pyrrolopyridinyl, azepinyl, diazepinyl, benzazepinyl, and the like.Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl,phenanthrolinyl, dibenzofuranyl, acridinyl, phenanthridinyl, xanthenyland the like.

The term “heteroarylalkyl” as used herein alone or as part of anothergroup refers to alkyl groups as defined above having a heteroarylsubstituent.

The terms “heterocycloalkyl” and, interchangeably, “heterocycle,” asused herein, alone or in combination, each refer to a saturated,partially unsaturated, or fully unsaturated monocyclic, bicyclic, ortricyclic heterocyclic group containing at least one heteroatom as aring member, wherein each said heteroatom may be independently chosenfrom nitrogen, oxygen, and sulfur. In certain embodiments, saidheterocycloalkyl will comprise from 1 to 4 heteroatoms as ring members.In further embodiments, said heterocycloalkyl will comprise from 1 to 2heteroatoms as ring members. In certain embodiments, saidheterocycloalkyl will comprise from 3 to 8 ring members in each ring. Infurther embodiments, said heterocycloalkyl will comprise from 3 to 7ring members in each ring. In yet further embodiments, saidheterocycloalkyl will comprise from 5 to 6 ring members in each ring.“Heterocycloalkyl” and “heterocycle” are intended to include sulfones,sulfoxides, N-oxides of tertiary nitrogen ring members, and carbocyclicfused and benzo fused ring systems; additionally, both terms alsoinclude systems where a heterocycle ring is fused to an aryl group, asdefined herein, or an additional heterocycle group. Examples ofheterocycle groups include aziridinyl, azetidinyl, 1,3-benzodioxolyl,dihydroisoindolyl, dihydroisoquinolinyl, dihydrocinnolinyl,dihydrobenzodioxinyl, dihydro[1,3]oxazolo[4,5-b]pyridinyl,benzothiazolyl, dihydroindolyl, dihy-dropyridinyl, 1,3-dioxanyl,1,4-dioxanyl, 1,3-dioxolanyl, imidazolidinyl, isoindolinyl, morpholinyl,oxazolidinyl, isoxazolidinyl, piperidinyl, piperazinyl,methylpiperazinyl, N-methylpiperazinyl, pyrrolidinyl, pyrazolidinyl,tetrahydrofuranyl, tetrahydropyridinyl, thiomorpholinyl, thiazolidinyl,diazepanyl, and the like. The heterocycle groups may be optionallysubstituted unless specifically prohibited.

The term “hydrazinyl” as used herein, alone or in combination, refers totwo amino groups joined by a single bond, i.e., —N—N—.

The term “hydroxy,” as used herein, alone or in combination, refers to—OH.

The term “hydroxyalkyl,” as used herein, alone or in combination, refersto a hydroxy group attached to the parent molecular moiety through analkyl group.

The term “hydroxamic acid”, as used herein, alone or in combination,refers to —C(═O)NHOH, wherein the parent molecular moiety is attached tothe hydroxamic acid group by means of the carbon atom.

The phrase “in the main chain” refers to the longest contiguous oradjacent chain of carbon atoms starting at the point of attachment of agroup to the compounds of any one of the formulas disclosed herein. Thephrase “linear chain of atoms” refers to the longest straight chain ofatoms independently selected from carbon, nitrogen, oxygen and sulfur.

The term “lower,” as used herein, alone or in a combination, where nototherwise specifically defined, means containing from 1 to and including6 carbon atoms.

The term “lower aryl,” as used herein, alone or in combination, meansphenyl or naphthyl, which may be optionally substituted as provided.

The term “lower heteroaryl,” as used herein, alone or in combination,means either 1) monocyclic heteroaryl comprising five or six ringmembers, of which between one and four said members may be heteroatomschosen from O, S, and N, or 2) bicyclic heteroaryl, wherein each of thefused rings comprises five or six ring members, comprising between themone to four heteroatoms chosen from O, S, and N.

The term “lower cycloalkyl,” as used herein, alone or in combination,means a monocyclic cycloalkyl having between three and six ring members.Lower cycloalkyls may be unsaturated. Examples of lower cycloalkylinclude cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “lower heterocycloalkyl,” as used herein, alone or incombination, means a monocyclic heterocycloalkyl having between threeand six ring members, of which between one and four may be heteroatomschosen from O, S, and N. Examples of lower heterocycloalkyls includepyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl,and morpholinyl. Lower heterocycloalkyls may be unsaturated.

The term “lower amino,” as used herein, alone or in combination, refersto —NRR, wherein R and R′ are independently chosen from hydrogen, loweralkyl, and lower heteroalkyl, any of which may be optionallysubstituted. Additionally, the R and R′ of a lower amino group maycombine to form a five- or six-membered heterocycloalkyl, either ofwhich may be optionally substituted.

The term “nitro,” as used herein, alone or in combination, refers to—NO₂.

The terms “oxy” or “oxa,” as used herein, alone or in combination, referto —O—.

The term “oxo,” as used herein, alone or in combination, refers to ═O.

The term “perhaloalkoxy” refers to an alkoxy group where all of thehydrogen atoms are replaced by halogen atoms.

The term “perhaloalkyl” as used herein, alone or in combination, refersto an alkyl group where all of the hydrogen atoms are replaced byhalogen atoms.

The term “phosphonate,” as used herein, alone or in combination, refersto a —P(═O)(OR)₂ group, wherein R is chosen from alkyl and aryl. Theterm “phosphonic acid”, as used herein, alone or in combination, refersto a —P(═O)(OH)₂ group.

The terms “sulfonate,” “sulfonic acid,” and “sulfonic,” as used herein,alone or in combination, refer to the —SO₃H group and its anion as thesulfonic acid is used in salt formation.

The term “sulfanyl,” as used herein, alone or in combination, refers to—S—.

The term “sulfinyl,” as used herein, alone or in combination, refers to—S(O)—.

The term “sulfonyl,” as used herein, alone or in combination, refers to—S(O)₂—.

The term “N-sulfonamido” refers to a RS(═O)₂NR′— group with R and R′ asdefined herein.

The term “S-sulfonamido” refers to a —S(═O)₂NRR′, group, with R and R′as defined herein.

The terms “thia” and “thio,” as used herein, alone or in combination,refer to a —S— group or an ether wherein the oxygen is replaced withsulfur. The oxidized derivatives of the thio group, namely sulfinyl andsulfonyl, are included in the definition of thia and thio.

The term “thiol,” as used herein, alone or in combination, refers to an—SH group.

The term “thiocarbonyl,” as used herein, when alone includes thioformyl—C(S)H and in combination is a —C(S)— group.

The term “N-thiocarbamyl” refers to an ROC(S)NR′— group, with R and R′as defined herein.

The term “O-thiocarbamyl” refers to a —OC(S)NRR′, group with R and R′ asdefined herein.

The term “thiocyanato” refers to a —CNS group.

The term “trihalomethoxy” refers to a X₃CO— group where X is a halogen.

Any definition herein may be used in combination with any otherdefinition to describe a composite structural group. By convention, thetrailing element of any such definition is that which attaches to theparent moiety. For example, the composite group alkylamido wouldrepresent an alkyl group attached to the parent molecule through anamido group, and the term alkoxyalkyl would represent an alkoxy groupattached to the parent molecule through an alkyl group.

When a group is defined to be “null,” what is meant is that said groupis absent. Similarly, when a designation such as “n” which may be chosenfrom a group or range of integers is designated to be 0, then the groupwhich it designates is either absent, if in a terminal position, orcondenses to form a bond, if it falls between two other groups.

The term “optionally substituted” means the anteceding group may besubstituted or unsubstituted. When substituted, the substituents of an“optionally substituted” group may include, without limitation, one ormore substituents independently selected from the following groups or aparticular designated set of groups, alone or in combination: loweralkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl,lower heterocycloalkyl, lower haloalkyl, lower haloalkenyl, lowerhaloalkynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl,phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, loweracyloxy, carbonyl, carboxyl, lower alkylcarbonyl, lower carboxyester,lower carboxamido, cyano, hydrogen, halogen, hydroxy, amino, loweralkylamino, arylamino, amido, nitro, thiol, lower alkylthio, lowerhaloalkylthio, lower perhaloalkylthio, arylthio, sulfonate, sulfonicacid, trisubstituted silyl, N₃, SH, SCH₃, C(O)CH₃, CO₂CH₃, CO₂H,pyridinyl, thiophene, furanyl, lower carbamate, and lower urea. Twosubstituents may be joined together to form a fused five-, six-, orseven-membered carbocyclic or heterocyclic ring consisting of zero tothree heteroatoms, for example forming methylenedioxy or ethylenedioxy.An optionally substituted group may be unsubstituted (e.g., —CH₂CH₃),fully substituted (e.g., —CF₂CF₃), monosubstituted (e.g., —CH₂CH₂F) orsubstituted at a level anywhere in-between fully substituted andmonosubstituted (e.g., —CH₂CF₃). Where substituents are recited withoutqualification as to substitution, both substituted and unsubstitutedforms are encompassed. Where a substituent is qualified as“substituted,” the substituted form is specifically intended.Additionally, different sets of optional substituents to a particularmoiety may be defined as needed; in these cases, the optionalsubstitution will be as defined, often immediately following the phrase,“optionally substituted with.”

When the construction

as used herein, the alkylene groups enclosed by ( )_(m) and ( )_(n) maybe m or n carbons long.

The term R or the term R′, appearing by itself and without a numberdesignation, unless otherwise defined, refers to a moiety chosen fromhydrogen, alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl andheterocycloalkyl, any of which may be optionally substituted. Such R andR′ groups should be understood to be optionally substituted as definedherein. Whether an R group has a number designation or not, every Rgroup, including R, R′ and R^(n) where n=(1, 2, 3, . . . n), everysubstituent, and every term should be understood to be independent ofevery other in terms of selection from a group. Should any variable,substituent, or term (e.g. aryl, heterocycle, R, etc.) occur more thanone time in a formula or generic structure, its definition at eachoccurrence is independent of the definition at every other occurrence.Those of skill in the art will further recognize that certain groups maybe attached to a parent molecule or may occupy a position in a chain ofelements from either end as written. Thus, by way of example only, anunsymmetrical group such as —C(O)N(R)— may be attached to the parentmoiety at either the carbon or the nitrogen.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as d-isomers and 1-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Starting compounds ofparticular stereochemistry are either commercially available or can bemade and resolved by techniques known in the art. Additionally, thecompounds disclosed herein may exist as geometric isomers. The presentinvention includes all cis, trans, syn, anti, entgegen (E), and zusammen(Z) isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Additionally, the compounds disclosed herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The term “bond” refers to a covalent linkage between two atoms, or twomoieties when the atoms joined by the bond are considered to be part oflarger substructure. A bond may be single, double, or triple unlessotherwise specified. A dashed line between two atoms in a drawing of amolecule indicates that an additional bond may be present or absent atthat position.

The term “disease” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disorder” and“condition” (as in medical condition), in that all reflect an abnormalcondition of the human or animal body or of one of its parts thatimpairs normal functioning, is typically manifested by distinguishingsigns and symptoms, and causes the human or animal to have a reducedduration or quality of life.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a therapeutic condition or disorderdescribed in the present disclosure. Such administration encompassesco-administration of these therapeutic agents in a substantiallysimultaneous manner, such as in a single capsule having a fixed ratio ofactive ingredients or in multiple, separate capsules for each activeingredient. In addition, such administration also encompasses use ofeach type of therapeutic agent in a sequential manner. In either case,the treatment regimen will provide beneficial effects of the drugcombination in treating the conditions or disorders described herein.

The phrase “therapeutically effective” is intended to qualify the amountof active ingredients used in the treatment of a disease or disorder.This amount will achieve the goal of reducing or eliminating the saiddisease or disorder.

The term “therapeutically acceptable” refers to those compounds (orsalts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitablefor use in contact with the tissues of patients without undue toxicity,irritation, and allergic response, are commensurate with a reasonablebenefit/risk ratio, and are effective for their intended use.

As used herein, reference to “treatment” of a patient is intended toinclude prophylaxis. The term “patient” means all mammals includinghumans. Examples of patients include humans, cows, dogs, cats, goats,sheep, pigs, and rabbits. Preferably, the patient is a human.

The term “prodrug” refers to a compound that is made more active invivo. Certain compounds disclosed herein may also exist as prodrugs, asdescribed in Hydrolysis in Drug and Prodrug Metabolism: Chemistry,Biochemistry, and Enzymology (Testa, Bernard and Mayer, Joachim M.Wiley-VHCA, Zurich, Switzerland 2003). Prodrugs of the compoundsdescribed herein are structurally modified forms of the compound thatreadily undergo chemical changes under physiological conditions toprovide the compound. Additionally, prodrugs can be converted to thecompound by chemical or biochemical methods in an ex vivo environment.For example, prodrugs can be slowly converted to a compound when placedin a transdermal patch reservoir with a suitable enzyme or chemicalreagent. Prodrugs are often useful because, in some situations, they maybe easier to administer than the compound, or parent drug. They may, forinstance, be bioavailable by oral administration whereas the parent drugis not. The prodrug may also have improved solubility in pharmaceuticalcompositions over the parent drug. A wide variety of prodrug derivativesare known in the art, such as those that rely on hydrolytic cleavage oroxidative activation of the prodrug. An example, without limitation, ofa prodrug would be a compound which is administered as an ester (the“prodrug”), but then is metabolically hydrolyzed to the carboxylic acid,the active entity. Additional examples include peptidyl derivatives of acompound.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The present invention includes compounds listed above in the formof salts, including acid addition salts. Suitable salts include thoseformed with both organic and inorganic acids. Such acid addition saltswill normally be pharmaceutically acceptable. However, salts ofnon-pharmaceutically acceptable salts may be of utility in thepreparation and purification of the compound in question. Basic additionsalts may also be formed and be pharmaceutically acceptable. For a morecomplete discussion of the preparation and selection of salts, refer toPharmaceutical Salts: Properties, Selection, and Use (Stahl, P.Heinrich. Wiley-VCHA, Zurich, Switzerland, 2002).

The term “therapeutically acceptable salt,” as used herein, representssalts or zwitterionic forms of the compounds disclosed herein which arewater or oil-soluble or dispersible and therapeutically acceptable asdefined herein. The salts can be prepared during the final isolation andpurification of the compounds or separately by reacting the appropriatecompound in the form of the free base with a suitable acid.Representative acid addition salts include acetate, adipate, alginate,L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate),bisulfate, butyrate, camphorate, camphorsulfonate, citrate, digluconate,formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate),lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate,3-phenylproprionate, phosphonate, picrate, pivalate, propionate,pyroglutamate, succinate, sulfonate, tartrate, L-tartrate,trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate,para-toluenesulfonate (p-tosylate), and undecanoate. Also, basic groupsin the compounds disclosed herein can be quaternized with methyl, ethyl,propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl,dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and sterylchlorides, bromides, and iodides; and benzyl and phenethyl bromides.Examples of acids which can be employed to form therapeuticallyacceptable addition salts include inorganic acids such as hydrochloric,hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic,maleic, succinic, and citric. Salts can also be formed by coordinationof the compounds with an alkali metal or alkaline earth ion. Hence, thepresent invention contemplates sodium, potassium, magnesium, and calciumsalts of the compounds disclosed herein, and the like.

Basic addition salts can be prepared during the final isolation andpurification of the compounds by reaction of a carboxy group with asuitable base such as the hydroxide, carbonate, or bicarbonate of ametal cation or with ammonia or an organic primary, secondary, ortertiary amine. The cations of therapeutically acceptable salts includelithium, sodium, potassium, calcium, magnesium, and aluminum, as well asnontoxic quaternary amine cations such as ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, diethylamine, ethylamine, tributylamine, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine,1-ephenamine, and N,N′-dibenzylethylenediamine. Other representativeorganic amines useful for the formation of base addition salts includeethylenediamine, ethanolamine, diethanolamine, piperidine, andpiperazine.

A salt of a compound can be made by reaction of the appropriatecompound, in the form of the free base, with the appropriate acid.

The compounds disclosed herein can exist as polymorphs and otherdistinct solid forms such as solvates, hydrates, and the like. Acompound may be a polymorph, solvate, or hydrate of a salt or of thefree base or acid.

While it may be possible for the compounds disclosed herein to beadministered as the raw chemical, it is also possible to present them aspharmaceutical formulations (equivalently, “pharmaceuticalcompositions”). Accordingly, provided herein are pharmaceuticalformulations which comprise one or more of certain compounds disclosedherein, or one or more pharmaceutically acceptable salts, esters,prodrugs, amides, or solvates thereof, together with one or morepharmaceutically acceptable carriers thereof and optionally one or moreother therapeutic ingredients. The carrier(s) must be “acceptable” inthe sense of being compatible with the other ingredients of theformulation and not deleterious to the recipient thereof. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, carriers, and excipients may be used assuitable and as understood in the art; e.g., in Remington'sPharmaceutical Sciences. The pharmaceutical compositions disclosedherein may be manufactured in any manner known in the art, e.g., bymeans of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

The formulations include those suitable for oral, parenteral (includingsubcutaneous, intradermal, intramuscular, intravenous, intraarticular,intraadiposal, intraarterial, intracranial, intralesional, intranasal,intraocular, intrapericardial, intraperitoneal, intrapleural,intraprostatical, intrarectal, intrathecal, intratracheal, intratumoral,intraumbilical, intravaginal, intravesicular, intravitreal, andintramedullary), intraperitoneal, rectal, topical (including, withoutlimitation, dermal, buccal, sublingual, vaginal, rectal, nasal, otic,and ocular), local, mucosal, sublingual, subcutaneous, transmucosal,transdermal, transbuccal, transdermal, and vaginal; liposomal, incremes, in lipid compositions, via a catheter, via a lavage, viacontinuous infusion, via infusion, via inhalation, via injection, vialocal delivery, via localized perfusion, bathing target cells directly,or any combination thereof. Administration although the most suitableroute may depend upon for example the condition and disorder of therecipient. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Typically, these methods include the step of bringing intoassociation a compound disclosed herein or a pharmaceutically acceptablesalt, ester, amide, prodrug or solvate thereof (“active ingredient”)with the carrier which constitutes one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers orfinely divided solid carriers or both and then, if necessary, shapingthe product into the desired formulation.

Formulations of the compounds disclosed herein suitable for oraladministration may be presented as discrete units such as hard or softcapsules, wafers, cachets or tablets each containing a predeterminedamount of the active ingredient; as a powder or granules; as a syrup,elixir, solution, or a suspension in an aqueous liquid or a non-aqueousliquid; or as an oil-in-water liquid emulsion, a water-in-oil liquidemulsion, or a compound dispersed in a liposome. The active ingredientmay also be presented as a bolus, electuary or paste.

Pharmaceutical preparations that can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated to provide delayed, slowed, orcontrolled release or absorption of the active ingredient therein.Compositions may further comprise an agent that enhances solubility ordispersability. All formulations for oral administration should be indosages suitable for such administration. The push-fit capsules cancontain the active ingredients in admixture with filler such as lactose,binders such as starches, and/or lubricants such as talc or magnesiumstearate and, optionally, stabilizers. In soft capsules, the activecompounds may be dissolved or suspended in suitable liquids, such asfatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added. Dragee cores are provided withsuitable coatings. For this purpose, concentrated sugar solutions may beused, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide,lacquer solutions, and suitable organic solvents or solvent mixtures.Dyestuffs or pigments may be added to the tablets or dragee coatings foridentification or to characterize different combinations of activecompound doses.

Depending on the route of administration, the compounds, or granules orparticles thereof, may be coated in a material to protect the compoundsfrom the action of acids and other natural conditions that mayinactivate the compounds.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion, either tothe body or to the site of a disease or wound. Formulations forinjection may be presented in unit dosage form, e.g., in ampoules or inmulti-dose containers, with an added preservative. The compositions maytake such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. The formulations may be presentedin unit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in powder form or in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or sterile pyrogen-free water,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Formulations for parenteral administration include aqueous andnon-aqueous (oily) sterile injection solutions of the active compoundswhich may contain antioxidants, buffers, bacteriostats and solutes whichrender the formulation isotonic with the blood of the intendedrecipient; and aqueous and non-aqueous sterile suspensions which mayinclude suspending agents and thickening agents. Suitable lipophilicsolvents or vehicles include fatty oils such as sesame oil, or syntheticfatty acid esters, such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions may contain substances that increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents that increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.To administer the therapeutic compound by other than parenteraladministration, it may be necessary to coat the compound with, orco-administer the compound with a material to prevent its inactivation(for example, via liposomal formulation).

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter, polyethylene glycol, or otherglycerides.

Certain compounds disclosed herein may be administered topically, thatis by non-systemic administration. This includes the application of acompound disclosed herein externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as gels, liniments, lotions, creams,ointments or pastes, and drops suitable for administration to the eye,ear or nose. The active ingredient for topical administration maycomprise, for example, from 0.001% to 10% w/w (by weight) of theformulation. In certain embodiments, the active ingredient may compriseas much as 10% w/w. In other embodiments, it may comprise less than 5%w/w. In certain embodiments, the active ingredient may comprise from 2%w/w to 5% w/w. In other embodiments, it may comprise from 0.1% to 1% w/wof the formulation.

Topical ophthalmic, otic, and nasal formulations disclosed herein maycomprise excipients in addition to the active ingredient. Excipientscommonly used in such formulations include, but are not limited to,tonicity agents, preservatives, chelating agents, buffering agents, andsurfactants. Other excipients comprise solubilizing agents, stabilizingagents, comfort-enhancing agents, polymers, emollients, pH-adjustingagents and/or lubricants. Any of a variety of excipients may be used informulations disclosed herein including water, mixtures of water andwater-miscible solvents, such as C1-C7-alkanols, vegetable oils ormineral oils comprising from 0.5 to 5% non-toxic water-soluble polymers,natural products, such as alginates, pectins, tragacanth, karaya gum,guar gum, xanthan gum, carrageenan, agar and acacia, starch derivatives,such as starch acetate and hydroxypropyl starch, and also othersynthetic products such as polyvinyl alcohol, polyvinylpyrrolidone,polyvinyl methyl ether, polyethylene oxide, preferably cross-linkedpolyacrylic acid and mixtures of those products. The concentration ofthe excipient is, typically, from 1 to 100,000 times the concentrationof the active ingredient. In preferred embodiments, the excipients to beincluded in the formulations are typically selected because of theirinertness towards the active ingredient component of the formulations.

Relative to ophthalmic, otic, and nasal formulations, suitabletonicity-adjusting agents include, but are not limited to, mannitol,sodium chloride, glycerin, sorbitol and the like. Suitable bufferingagents include, but are not limited to, phosphates, borates, acetatesand the like. Suitable surfactants include, but are not limited to,ionic and nonionic surfactants (though nonionic surfactants arepreferred), RLM 100, POE 20 cetylstearyl ethers such as Procol® CS20 andpoloxamers such as Pluronic® F68.

The formulations set forth herein may comprise one or morepreservatives. Examples of such preservatives include p-hydroxybenzoicacid ester, sodium perborate, sodium chlorite, alcohols such aschlorobutanol, benzyl alcohol or phenyl ethanol, guanidine derivativessuch as polyhexamethylene biguanide, sodium perborate,polyquarternium-1, amino alcohols such as AMP-95, or sorbic acid. Incertain embodiments, the formulation may be self-preserved so that nopreservation agent is required.

In certain topical embodiments, formulations are prepared using abuffering system that maintains the formulation at a pH of about 4.5 toa pH of about 8. In further embodiments, the pH is from 7 to 8.

Gels for topical or transdermal administration may comprise, generally,a mixture of volatile solvents, nonvolatile solvents, and water. Incertain embodiments, the volatile solvent component of the bufferedsolvent system may include lower (C1-C6) alkyl alcohols, lower alkylglycols and lower glycol polymers. In further embodiments, the volatilesolvent is ethanol. The volatile solvent component is thought to act asa penetration enhancer, while also producing a cooling effect on theskin as it evaporates. The nonvolatile solvent portion of the bufferedsolvent system is selected from lower alkylene glycols and lower glycolpolymers. In certain embodiments, propylene glycol is used. Thenonvolatile solvent slows the evaporation of the volatile solvent andreduces the vapor pressure of the buffered solvent system. The amount ofthis nonvolatile solvent component, as with the volatile solvent, isdetermined by the pharmaceutical compound or drug being used. When toolittle of the nonvolatile solvent is in the system, the pharmaceuticalcompound may crystallize due to evaporation of volatile solvent, whilean excess may result in a lack of bioavailability due to poor release ofdrug from solvent mixture. The buffer component of the buffered solventsystem may be selected from any buffer commonly used in the art; incertain embodiments, water is used. A common ratio of ingredients isabout 20% of the nonvolatile solvent, about 40% of the volatile solvent,and about 40% water. Several optional ingredients can be added to thetopical composition. These include, but are not limited to, chelatorsand gelling agents. Appropriate gelling agents can include, but are notlimited to, semisynthetic cellulose derivatives (such ashydroxypropylmethylcellulose) and synthetic polymers, galactomannanpolymers (such as guar and derivatives thereof), and cosmetic agents.

Lotions include those suitable for application to the skin or eye. Aneye lotion may comprise a sterile aqueous solution optionally containinga bactericide and may be prepared by methods similar to those for thepreparation of drops. Lotions or liniments for application to the skinmay also include an agent to hasten drying and to cool the skin, such asan alcohol or acetone, and/or a moisturizer such as glycerol or an oilsuch as castor oil or arachis oil.

Creams, ointments or pastes are semi-solid formulations of the activeingredient for external application. They may be made by mixing theactive ingredient in finely-divided or powdered form, alone or insolution or suspension in an aqueous or non-aqueous fluid, with the aidof suitable machinery, with a greasy or non-greasy base. The base maycomprise hydrocarbons such as hard, soft or liquid paraffin, glycerol,beeswax, a metallic soap; a mucilage; an oil of natural origin such asalmond, corn, arachis, castor or olive oil; wool fat or its derivativesor a fatty acid such as stearic or oleic acid together with an alcoholsuch as propylene glycol or a macrogel. The formulation may incorporateany suitable surface active agent such as an anionic, cationic ornon-ionic surfactant such as a sorbitan ester or a polyoxyethylenederivative thereof. Suspending agents such as natural gums, cellulosederivatives or inorganic materials such as silicaceous silicas, andother ingredients such as lanolin, may also be included.

Drops may comprise sterile aqueous or oily solutions or suspensions andmay be prepared by dissolving the active ingredient in a suitableaqueous solution of a bactericidal and/or fungicidal agent and/or anyother suitable preservative, and, in certain embodiments, including asurface active agent. The resulting solution may then be clarified byfiltration, transferred to a suitable container which is then sealed andsterilized by autoclaving or maintaining at 98-100° C. for half an hour.Alternatively, the solution may be sterilized by filtration andtransferred to the container by an aseptic technique. Examples ofbactericidal and fungicidal agents suitable for inclusion in the dropsare phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride(0.01%) and chlorhexidine acetate (0.01%). Suitable solvents for thepreparation of an oily solution include glycerol, diluted alcohol andpropylene glycol.

Formulations for topical administration in the mouth, for examplebuccally or sublingually, include lozenges comprising the activeingredient in a flavored basis such as sucrose and acacia or tragacanth,and pastilles comprising the active ingredient in a basis such asgelatin and glycerin or sucrose and acacia.

For administration by inhalation, compounds may be convenientlydelivered from an insufflator, nebulizer pressurized packs or otherconvenient means of delivering an aerosol spray. Pressurized packs maycomprise a suitable propellant such as dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol, the dosageunit may be determined by providing a valve to deliver a metered amount.Alternatively, for administration by inhalation or insufflation, thecompounds according to the invention may take the form of a dry powdercomposition, for example, a powder mix of the compound and a suitablepowder base such as lactose or starch. The powder composition may bepresented in unit dosage form, in for example, capsules, cartridges,gelatin or blister packs from which the powder may be administered withthe aid of an inhalator or insufflator.

The therapeutic compound may also be administered intraspinally orintracerebrally. Dispersions for these types of administrations can beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations may contain a preservative to prevent the growth ofmicroorganisms.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. In all cases, the composition must be sterileand must be fluid to the extent that easy syringability exists. It mustbe stable under the conditions of manufacture and storage and must bepreserved against the contaminating action of microorganisms such asbacteria and fungi. The carrier can be a solvent or dispersion mediumcontaining, for example, water, ethanol, polyol (such as, glycerol,propylene glycol, and liquid polyethylene glycol, and the like),suitable mixtures thereof, and vegetable oils. The proper fluidity canbe maintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case of dispersionand by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, sodium chloride, orpolyalcohols such as mannitol and sorbitol, in the composition.

Sterile injectable solutions can be prepared by incorporating thetherapeutic compound in the required amount in an appropriate solventwith one or a combination of ingredients enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the therapeutic compound into a sterile carrier thatcontains a basic dispersion medium and required other ingredients to bepharmacologically sound. In the case of sterile powders for thepreparation of sterile injectable solutions, the preferred methods ofpreparation are vacuum drying and freeze-drying which yields a powder ofthe active ingredient (i.e., the therapeutic compound) plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

It is especially advantageous to formulate parenteral compositions indosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the subjects to be treated; each unitcontaining a predetermined quantity of therapeutic compound calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. The specification for the dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding such a therapeutic compound for the treatment ofa selected condition in a patient.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations described above may include otheragents conventional in the art having regard to the type of formulationin question, for example, those suitable for oral administration mayinclude flavoring agents.

Compounds may be administered at a dose of from 0.1 to 500 mg/kg perday. The dose range for adult humans is generally from 5 mg to 2 g/day.Tablets or other forms of presentation provided in discrete units mayconveniently contain an amount of one or more compounds which iseffective at such dosage or as a multiple of the same, for instance,units containing 5 mg to 500 mg, usually around 10 mg to 200 mg.

Preferred unit dosage formulations are those containing an effectivedose, as herein below recited, or an appropriate fraction thereof, ofthe active ingredient. In certain embodiments, a formulation disclosedherein is administered once a day. However, the formulations may also beformulated for administration at any frequency of administration,including once a week, once every 5 days, once every 3 days, once every2 days, twice a day, three times a day, four times a day, five times aday, six times a day, eight times a day, every hour, or any greaterfrequency. Such dosing frequency is also maintained for a varyingduration of time depending on the therapeutic regimen. The duration of aparticular therapeutic regimen may vary from one-time dosing to aregimen that extends for months or years. The formulations areadministered at varying dosages, but typical dosages are one to twodrops at each administration, or a comparable amount of a gel or otherformulation. One of ordinary skill in the art would be familiar withdetermining a therapeutic regimen for a specific indication.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. Similarly, theprecise amount of compound administered to a patient will be theresponsibility of the attendant physician. The specific dose level forany particular patient will depend upon a variety of factors includingthe activity of the specific compound employed, the age, body weight,general health, sex, diets, time of administration, route ofadministration, rate of excretion, drug combination, the precisedisorder being treated, and the severity of the indication or conditionbeing treated. In addition, the route of administration may varydepending on the condition and its severity.

In certain instances, it may be appropriate to administer at least oneof the compounds described herein (or a pharmaceutically acceptablesalt, ester, or prodrug thereof) in combination with another therapeuticagent. By way of example only, if one of the side effects experienced bya patient upon receiving one of the compounds herein is inflammation,then it may be appropriate to administer an anti-inflammatory agent incombination with the initial therapeutic agent. Alternatively, by way ofexample only, the therapeutic effectiveness of one of the compoundsdescribed herein may be enhanced by administration of an adjuvant (i.e.,by itself the adjuvant may only have minimal therapeutic benefit, but incombination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). There is even the possibility thattwo compounds, one of the compounds described herein and a secondcompound may together achieve the desired therapeutic effect thatneither alone could achieve. Alternatively, by way of example only, thebenefit experienced by a patient may be increased by administering oneof the compounds described herein with another therapeutic agent (whichalso includes a therapeutic regimen) that also has therapeutic benefit.By way of example only, in a treatment for acute myelogenous leukemia orsickle cell anemia involving administration of one of the compoundsdescribed herein, increased therapeutic benefit may result by alsoproviding the patient with another therapeutic agent for sickle cellanemia or for acute myelogenous leukemia. In any case, regardless of thedisease, disorder or condition being treated, the overall benefitexperienced by the patient may simply be additive of the two therapeuticagents or the two agents may have synergistic therapeutic effects in apatient.

Effective combination therapy may be achieved with a single compositionor pharmacological formulation that includes both agents, or with twodistinct compositions or formulations, at the same time, wherein onecomposition includes a compound of the present disclosure, and the otherincludes the second agent(s). Alternatively, the therapy may precede orfollow the other agent treatment by intervals ranging from minutes tomonths. Administration of the compounds of the present disclosure to apatient will follow general protocols for the administration ofpharmaceuticals, taking into account the toxicity, if any, of the drug.It is expected that the treatment cycles would be repeated as necessary.

Specific, non-limiting examples of possible combination therapiesinclude use of certain compounds of the invention with the followingagents and classes of agents: agents that inhibit DNA methyltransferasessuch as decitabine or 5′-aza-cytadine; agents that inhibit the activityof histone deacetylases, histone de-sumoylases, histonede-ubiquitinases, or histone phosphatases such as hydroxyurea; antisenseRNAs that might inhibit the expression of other components of theprotein complex bound at the DR site in the gamma globin promoter;agents that inhibit the action of Klf1 or the expression of KLF1; agentsthat inhibit the action of Bcl11a or the expression of BCL11A; andagents that inhibit cell cycle progression such as hydroxyurea, ara-C ordaunorubicin; agents that induce differentiation in leukemic cells suchas all-trans retinoic acid (ATRA).

Thus, in another aspect, the present invention provides methods fortreating diseases or disorders in a human or animal subject in need ofsuch treatment comprising administering to said subject an amount of acompound disclosed herein effective to reduce or prevent said disorderin the subject, optionally in combination with at least one additionalagent for the treatment of said disorder that is known in the art.

Used either as a monotherapy or in combination with other agents, thecompounds disclosed herein are useful in the prevention and/or treatmentof beta-hemoglobinopathies such as thalassemia major, sickle celldisease, hemoglobin E/thalassemia, and thalassemia intermedia.

The compounds disclosed herein can be used in the treatment of diseasesin which an increase in transcription through the manipulation ofepigenetic regulatory factors such as inhibition of KDM1A would bebeneficial to the patient. This applies to diseases including but notlimited to loss of function mutations, mutations resulting inhaploinsufficiency, deletions and duplications of genetic material orepigenetic regulatory mechanisms have altered the normal expressionpattern of a gene or genes that has the effect of altering the dose of agene product(s). Such diseases may include diseases both acquired andhereditary in which the expression of, for example, cytokines affectingimmune function, are altered, X-linked mental retardation and otherforms of compromised cognitive or motor function such as Alzheimer andParkinson disease whether they are the acquired or hereditary forms,lipid disorders such as elevated cholesterol, low density lipoprotein,very low density lipoprotein or triglycerides, both type one and typetwo diabetes, and Mendelian genetic diseases.

Other disorders or conditions that can be advantageously treated by thecompounds disclosed herein include inflammation and inflammatoryconditions. Inflammatory conditions include, without limitation:arthritis, including sub-types and related conditions such as rheumatoidarthritis, spondyloarthropathies, gouty arthritis, osteoarthritis,systemic lupus erythematosus, juvenile arthritis, acute rheumaticarthritis, enteropathic arthritis, neuropathic arthritis, psoriaticarthritis, and pyogenic arthritis; osteoporosis, tendonitis, bursitis,and other related bone and joint disorders; gastrointestinal conditionssuch as reflux esophagitis, diarrhea, inflammatory bowel disease,Crohn's disease, gastritis, irritable bowel syndrome, ulcerativecolitis, acute and chronic inflammation of the pancreas; pulmonaryinflammation, such as that associated with viral infections and cysticfibrosis; skin-related conditions such as psoriasis, eczema, burns,sunburn, dermatitis (such as contact dermatitis, atopic dermatitis, andallergic dermatitis), and hives; pancreatitis, hepatitis, pruritus andvitiligo. In addition, compounds of invention are also useful in organtransplant patients either alone or in combination with conventionalimmunomodulators.

Autoimmune disorders may be ameliorated by the treatment with compoundsdisclosed herein. Autoimmune disorders include Crohn's disease,ulcerative colitis, dermatitis, dermatomyositis, diabetes mellitus type1, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome(GBS), autoimmune encephalomyelitis, Hashimoto's disease, idiopathicthrombocytopenic purpura, lupus erythematosus, mixed connective tissuedisease, multiple sclerosis (MS), myasthenia gravis, narcolepsy,pemphigus vulgaris, pernicious anemia, psoriasis, psoriatic arthritis,polymyositis, primary biliary cirrhosis, rheumatoid arthritis, Sjögren'ssyndrome, scleroderma, temporal arteritis (also known as “giant cellarteritis”), vasculitis, and Wegener's granulomatosis.

The compounds disclosed herein are also useful for the treatment oforgan and tissue injury associated with severe burns, sepsis, trauma,wounds, and hemorrhage- or resuscitation-induced hypotension, and alsoin such diseases as vascular diseases, migraine headaches, periarteritisnodosa, thyroiditis, aplastic anemia, Hodgkin's disease, scleredoma,rheumatic fever, type I diabetes, neuromuscular junction diseaseincluding myasthenia gravis, white matter disease including multiplesclerosis, sarcoidosis, nephritis, nephrotic syndrome, Behcet'ssyndrome, polymyositis, gingivitis, periodontis, swelling occurringafter injury, ischemias including myocardial ischemia, cardiovascularischemia, and ischemia secondary to cardiac arrest, and the like.

The compounds disclosed herein are also useful for the treatment ofcertain diseases and disorders of the nervous system. Central nervoussystem disorders in KDM1A inhibition is useful include corticaldementias including Alzheimer's disease, central nervous system damageresulting from stroke, ischemias including cerebral ischemia (both focalischemia, thrombotic stroke and global ischemia (for example, secondaryto cardiac arrest), and trauma. Neurodegenerative disorders in whichKDM1A inhibition is useful include nerve degeneration or nerve necrosisin disorders such as hypoxia, hypoglycemia, epilepsy, and in cases ofcentral nervous system (CNS) trauma (such as spinal cord and headinjury), hyperbaric oxygen-induced convulsions and toxicity, dementiae.g., pre-senile dementia, and AIDS-related dementia, cachexia,Sydenham's chorea, Huntington's disease, Parkinson's Disease,amyotrophic lateral sclerosis (ALS), Korsakoff's disease, cognitivedisorders relating to a cerebral vessel disorder, hypersensitivity,sleeping disorders, schizophrenia, depression, depression or othersymptoms associated with Premenstrual Syndrome (PMS), and anxiety.

Still other disorders or conditions advantageously treated by thecompounds disclosed herein include the prevention or treatment ofhyperproliferative diseases, especially cancers, either alone or incombination with standards of care especially those agents that targettumor growth by re-instating tumor suppressor genes in the malignantcells. Hematological and non-hematological malignancies which may betreated or prevented include but are not limited to multiple myeloma,acute and chronic leukemias and hematopoietic proliferative andneoplastic disorders including Myelodysplastic Syndrome (MDS), AcuteMyelogenous Leukemia (AML), Acute Lymphocytic Leukemia (ALL), ChronicLymphocytic Leukemia (CLL), and Chronic Myelogenous Leukemia (CML),lymphomas, including Hodgkin's lymphoma and non-Hodgkin's lymphoma (low,intermediate, and high grade), as well as solid tumors and malignanciesof the brain, head and neck, breast, lung (including non-small-cell lungcancer), reproductive tract, upper digestive tract, pancreas, liver,renal system, bladder, prostate and colorectal. The present compoundsand methods can also be used to treat fibrosis, such as that whichoccurs with radiation therapy. The present compounds and methods can beused to treat subjects having or prevent the progression of adenomatouspolyps, including those with familial adenomatous polyposis (FAP) orsarcoidosis. Non-cancerous proliferative disorders additionally includepsoriasis, eczema, and dermatitis.

The present compounds may also be used in co-therapies, partially orcompletely, in place of other conventional anti-inflammatory therapies,such as together with steroids, NSAIDs, COX-2 selective inhibitors,5-lipoxygenase inhibitors, LTB₄ antagonists and LTA₄ hydrolaseinhibitors. The compounds disclosed herein may also be used to preventtissue damage when therapeutically combined with antibacterial orantiviral agents.

The compounds disclosed herein are also useful for the treatment oftreat metabolic disorders. KDM1A, using flavin adenosine dinucleotide(FAD) as a cofactor, epigenetically regulates energy-expenditure genesin adipocytes depending on the cellular FAD availability. Additionally,loss of KDM1A function induces a number of regulators of energyexpenditure and mitochondrial metabolism resulting in the activation ofmitochondrial respiration. Furthermore, in the adipose tissues from micefed a high-fat diet, expression of KDM1A-target genes is reduced.

Metabolic syndrome (also known as metabolic syndrome X) is characterizedby having at least three of the following symptoms: insulin resistance;abdominal fat—in men this is defined as a 40 inch waist or larger, inwomen 35 inches or larger; high blood sugar levels—at least 110milligrams per deciliter (mg/dL) after fasting; high triglycerides—atleast 150 mg/dL in the blood stream; low HDL—less than 40 mg/dL;pro-thrombotic state (e.g., high fibrinogen or plasminogen activatorinhibitor in the blood); or blood pressure of 130/85 mmHg or higher. Aconnection has been found between metabolic syndrome and otherconditions such as obesity, high blood pressure and high levels of LDLcholesterol, all of which are risk factors for cardiovascular diseases.For example, an increased link between metabolic syndrome andatherosclerosis has been shown. People with metabolic syndrome are alsomore prone to developing type 2 diabetes, as well as PCOS (polycysticovarian syndrome) in women and prostate cancer in men.

As described above, insulin resistance can be manifested in severalways, including type 2 diabetes. Type 2 diabetes is the condition mostobviously linked to insulin resistance. Compensatory hyperinsulinemiahelps maintain normal glucose levels often for decades before overtdiabetes develops. Eventually the beta cells of the pancreas are unableto overcome insulin resistance through hypersecretion. Glucose levelsrise and a diagnosis of diabetes can be made. Patients with type 2diabetes remain hyperinsulinemic until they are in an advanced stage ofdisease. As described above, insulin resistance can also correlate withhypertension. One half of patients with essential hypertension areinsulin resistant and hyperinsulinemic, and there is evidence that bloodpressure is linked to the degree of insulin resistance. Hyperlipidemia,too, is associated with insulin resistance. The lipid profile ofpatients with type 2 diabetes includes increased serum very-low-densitylipoprotein (VLDL) cholesterol and triglyceride levels and, sometimes, adecreased low-density lipoprotein (LDL) cholesterol level. Insulinresistance has been found in persons with low levels of high-densitylipoprotein HDL). Insulin levels have also been linked to VLDL synthesisand plasma triglyceride levels.

Specific metabolic diseases and symptoms to be treated by the compounds,compositions, and methods disclosed herein are those mediated at leastin part by KDM1A. Accordingly, disclosed herein are methods: fortreating insulin resistance in a subject; for reducing glycogenaccumulation in a subject; for raising HDL or HDLc, lowering LDL orLDLc, shifting LDL particle size from small dense to normal LDL,lowering VLDL, lowering triglycerides, or inhibiting cholesterolabsorption in a subject; for reducing insulin resistance, enhancingglucose utilization or lowering blood pressure in a subject; forreducing visceral fat in a subject; for reducing serum transaminases ina subject; for inducing mitochondrial respiration in a subject; or fortreating disease; all comprising the administration of a therapeuticamount of a compound as described herein, to a patient in need thereof.In further embodiments, the disease to be treated may be a metabolicdisease. In further embodiment, the metabolic disease may be selectedfrom the group consisting of: obesity, diabetes mellitus, especiallyType 2 diabetes, hyperinsulinemia, glucose intolerance, metabolicsyndrome X, dyslipidemia, hypertriglyceridemia, hypercholesterolemia,and hepatic steatosis. In other embodiments, the disease to be treatedmay be selected from the group consisting of: cardiovascular diseasesincluding vascular disease, atherosclerosis, coronary heart disease,cerebrovascular disease, heart failure and peripheral vessel disease. Inpreferred embodiments, the methods above do not result in the inductionor maintenance of a hypoglycemic state.

Besides being useful for human treatment, certain compounds andformulations disclosed herein may also be useful for veterinarytreatment of companion animals, exotic animals and farm animals,including mammals, rodents, and the like. More preferred animals includehorses, dogs, and cats.

Methods

General Synthetic Methods for Preparing Compounds

The following invention is further illustrated by the followingExamples.

In the Examples below and throughout the disclosure, the followingabbreviations may be used: PTFE=polytetrafluoroethylene; RM=ReactionMixture; R H=Relative Humidity; RT=Room Temperature; SM=StartingMaterial; MeCN=acetonitrile; ClPh=chlorophenol; DCE=dichloroethane;DCM=dichloromethane; DIPE=di-isopropylether; DMA=dimethyl acetamide;DMF=dimethyl formamide; DMSO=dimethylsulfoxide; Et₂O=diethyl ether;EtOAc=ethyl acetate; EtOH=ethanol; H₂O=water; I PA=propan-2-ol;i-PrOAc=iso-propyl acetate; MEK=methyl ethyl ketone; MeOH=methanol;MIBK=methyl isobutyl ketone; MTBE=methyl tert-butyl ether;n-BuOAc=n-butyl acetate; n-BuOH=n-butanol; NMP=n-methyl pyrrolidone;n-PrOH=n-propanol; s-BuOAc=s-butyl acetate; t-BuOH=t-butanol;TFA=tri-fluoro acetic acid; THF=tetrahydrofuran;TMP=2,2,4-trimethylpentane; ¹H-NMR=Proton Nuclear magnetic Resonance;DSC=Differential Scanning calorimetry; DVS=Dynamic Vapour Sorption;GVS=Gravimetric Vapour Sorption; HPLC=High Performance LiquidChromatography; HS=Head Space; HSM=Hot Stage Microscopy; IC=IonChromatography; IDR=Intrinsic Dissolution Rate; KF=Karl-Fisher;MAS=Magic Angle Spinning; MDSC=Modulated Differential Scanningcalorimetry; PLM=Polarised Light Microscopy; PVM=Particle Vision andMeasurement; SCXRD=Single Crystal X-Ray Diffraction; SS-NMR=Solid StateNuclear Magnetic Resonance; TGA=Thermal Gravimetric Analysis;UV=UltraViolet VH-XRPD=Variable Humidity X-Ray Powder Diffraction;VT-XRPD=Variable Temperature X-Ray Powder Diffraction; and XRPD=X-RayPowder Diffraction. Other abbreviations may be used and will be familiarin context to those of skill in the art.

Example 1

Synthesis of 1

4-(1H-1,2,3-triazolyl-1-yl)benzoyl chloride (201)

In a 100-mL round-bottom flask were combined a solution of4-(1H-1,2,3-triazol-1-yl)benzoic acid (1.2 g, 6.34 mmol, 1.00 equiv) inthionyl chloride (20 mL). The resulting solution was stirred for 1 h at80° C., then concentrated under vacuum, affording 1.2 g (78%) of theproduct (as its hydrochloride salt) as an off-white solid.

1-(4-(1H-1,2,3-triazol-1-yl)benzoyl-2-carbomethoxy aziridine (203)

In a 250-mL round-bottom flask were added a solution of methylaziridine-2-carboxylate (1 g, 9.89 mmol, 1.00 equiv) in CH₂Cl₂ (80 mL)and Et₃N (3 g, 29.65 mmol, 3.00 equiv), followed by the addition of asolution of 4-(1H-1,2,3-triazol-1-yl)benzoyl chloride (2.3 g, 11.08mmol, 1.12 equiv) in CH₂Cl₂ (20 mL) dropwise with stirring at 0° C. Theresulting solution was stirred for 1 h at 25° C., then washed with 1×50mL of water and 1×50 mL of brine, dried over Na₂SO₄, and concentratedunder vacuum. The residue was applied onto a silica gel column withethyl acetate/petroleum ether (1:3). This resulted in 2.5 g (93%) of theproduct as a white solid.

Methyl3-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]propanoate

In a 50-mL round-bottom flask were combined the compound produced in theprevious step (1.5 g, 5.51 mmol, 1.00 equiv), CH₃CN (20 mL),(1S,2R)-2-(4-fluorophenyl)cyclopropan-1-amine hydrochloride (2.6 g,13.86 mmol, 2.50 equiv) and Et₃N (1.4 g, 13.84 mmol, 2.48 equiv). Theresulting solution was stirred for 16 h at 80° C. in an oil bath, thendiluted with 50 ml of EtOAc, washed with 1×30 mL of water and 1×30 mL ofbrine, dried over Na₂SO₄ and concentrated under vacuum. The residue wasapplied onto a silica gel column with EtOAc/petroleum ether (2:1),affording 1 g (43%) of PH-IMA-2013-003-362-11 as an off-white solid.

Methyl3-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](tert-butoxycarbonyl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]propanoate(204)

In a 50-mL round-bottom flask were combined the compound produced in theprevious step (1 g, 2.36 mmol, 1.00 equiv), CH₂Cl₂ (20 mL), Et₃N (500mg, 4.94 mmol, 2.09 equiv) and di-tert-butyl dicarbonate (780 mg, 3.57mmol, 1.51 equiv). The resulting solution was stirred for 16 h at 25° C.The resulting mixture was washed with 1×30 mL of water and 1×30 mL ofbrine, dried over Na₂SO₄, and concentrated under vacuum. The residue wasapplied onto a silica gel column with EtOAc/petroleum ether (1:2),affording 600 mg (49%) of PH-IMA-2013-003-362-12 as a white solid.

3-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](tert-butoxycarbonyl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]propanoicacid (205)

Into a 50-mL round-bottom flask was added a solution of the compoundproduced in the previous step (600 mg, 1.15 mmol, 1.00 equiv) in THF (25mL), followed by LiOH (41 mg, 1.71 mmol, 1.49 equiv) in water (6 mL).The resulting solution was stirred for 2 h at 25° C. The pH value of thesolution was adjusted to 5 with HCl (2 M). The resulting solution wasextracted with 2×30 mL of EtOAc, and the organic layers were combinedand washed with 1×30 mL of brine, dried over anhydrous Na₂SO₄, andconcentrated under vacuum, affording 580 mg (99%) of the product as awhite solid.

N-[3-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](tert-butoxycarbonyl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(206)

In a 100-mL round-bottom flask were combined the compound from theprevious step (580 mg, 1.14 mmol, 1.00 equiv), THF (40 mL), and3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (“DEPBT”) (530mg, 1.77 mmol, 1.56 equiv), followed by the addition of imidazole (120mg, 1.76 mmol, 1.55 equiv) at 0° C. The mixture was stirred for 40 min.To this was added a solution of 1-methylpiperazine (180 mg, 1.80 mmol,1.58 equiv) in THF (10 mL) dropwise with stirring at 0° C. The resultingsolution was stirred for 16 h at 25° C., diluted with 50 mL of EtOAc,then washed with 1×50 mL of sat. NaHCO₃ and 1×50 mL of brine. Theorganic layers were dried over Na₂SO₄ and concentrated under vacuum,affording 600 mg (89%) of the product as light yellow oil.

N-[3-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(1)

In a 50-mL round-bottom flask were combined the compound from theprevious step (600 mg, 1.01 mmol, 1.00 equiv), CH₂Cl₂ (20 mL) andCF₃COOH (4 mL). The resulting solution was stirred for 2 h at 25° C. ThepH value of the solution was adjusted to 9 with sat. NaHCO₃. Theresulting solution was extracted with 3×30 mL of EtOAc and the organiclayers were combined and washed with 1×50 mL of brine. The organiclayers was dried over Na₂SO₄ and concentrated under vacuum. The crudeproduct (5 mL) was purified by Prep-HPLC with the following conditions(2#-AnalyseHPLC-SHIMADZU (HPLC-10)): Column, XBridge C18 OBD PrepColumn, 100? 10 μm, 19 mm×250 mm; mobile phase, Waters (10 MMOL/LNH4HCO3) and ACN—Waters (20.0% ACN—Waters up to 60.0% in 6 min);Detector, uv 254/220 nm. 150 mL product was obtained. This resulted in330 mg (66%) of 1 as a white solid.

Example 4

Synthesis of 4

4-phenylbenzoyl chloride

Into a 250-mL round-bottom flask were combined 4-phenylbenzoic acid (15g, 75.67 mmol, 1.00 equiv) and thionyl chloride (30 mL). The resultingsolution was stirred for 16 h at 80° C. in an oil bath, thenconcentrated under vacuum, resulting in 15 g (91%) of the product as anoff-white solid.

(2S)-2-[(4-(phenyl)phenyl)formamido]-4-[tert-butyldiphenylsilyl)oxy]butanoicacid

The method used to prepare 203 was used with(2S)-2-amino-4-[(tert-butyldiphenylsilyl)oxy]butanoic acid (6.8 g, 19.02mmol, 1.00 equiv) and 4-phenylbenzoyl chloride (5 g, 23.08 mmol, 1.20equiv), affording 8 g (78%) of PH-IMA-2013-003-174-2 as a yellow oil.

N-[1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxo-4-((tert-butyldiphenylsilyl)oxy)-butan-2-yl]-4-phenylbenzamide

The method used to prepare 206 was used with the compound from theprevious step (5 g, 9.29 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide hydrochloride (2.4 g, 13.98 mmol, 1.50 equiv)to afford 5 g (83.3%) of the product as a off-white solid.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[4-(phenyl)phenyl)formamido]-1-butanol(207)

In a 250-mL round-bottom flask were combined the compound from theprevious step (22 g, 33.59 mmol, 1.00 equiv) and THF (150 ml). Bu₄NF (66mL, 66 mmol, 1.0 M in THF) was added dropwise with stirring at 0° C. Theresulting solution was stirred for 16 h at 25° C., concentrated undervacuum, diluted with 100 mL of EtOAc, and washed with 4×100 mL of waterand 2×100 mL of brine. The combined organic layers was dried over Na₂SO₄and applied onto a silica gel column with EtOAc to afford 10 g (71%) ofthe product as a off-white solid.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[4-(phenyl)phenyl)formamido]-1-butanal(208)

In a 500-mL flask purged and maintained with an inert atmosphere ofnitrogen were combined the compound produced in the previous step (10 g,24.01 mmol, 1.00 equiv) in CH₂Cl₂ (250 ml). This was followed by theaddition of Des s-Martin periodinane (20.4 g, 48.11 mmol, 2.00 equiv),in portions at 0° C. The resulting solution was stirred for 1 h at 25°C., then filtered out over diatomaceous earth, concentrated underreduced pressure, diluted with 30 mL CH₂Cl₂, and applied onto a silicagel column with EtOAc/petroleum ether (1:1) to afford 9 g (90%) of theproduct as a white solid.

N-[(2S)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl]-4-phenylbenzamide(4)

In a 50-mL flask purged and maintained with an inert atmosphere ofnitrogen was combined a solution of the compound from the previous step(450 mg, 1.09 mmol, 1.00 equiv), CH₂Cl₂ (10 ml), and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (30 mg, 0.39 mmol, 1.20equiv). This was followed by the addition of NaBH(AcO)₃ (552 mg, 2.60mmol, 2.40 equiv), in portions at 0° C. The resulting solution wasstirred for 10 min at 25° C. The reaction was then quenched by theaddition of 5 mL of water. The resulting solution was extracted with3×10 mL of CH₂Cl₂, and the organic layers were combined, dried overNa₂SO₄, concentrated under reduced pressure, and purified with Prep-HPLCto afford 223.9 mg (38%) of the product as a white solid.

Example 12

Synthesis of 12

4-(1H-Pyrazol-1-yl)benzoyl chloride

The method used to prepare 201 was used with 4-(1H-pyrazol-1-yl)benzoicacid (100 mg, 0.53 mmol), affording 110 mg (crude) of4-(1H-pyrazol-1-yl)benzoyl chloride as a yellow solid.

tert-ButylN-[(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[[4-(1H-pyrazol-1-yl)phenyl]formamido]butyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate

The method used to prepare 203 was used with tert-butylN-[(3S)-3-amino-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate(227 mg, 0.48 mmol, 1.00 equiv) and 4-(1H-pyrazol-1-yl)benzoyl chloride(110 mg, 0.53 mmol, 1.10 equiv), affording 150 mg (49%) of the productas a yellow oil.

N-[(2S)-4-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl]-4-(1H-pyrazol-1-yl)benzamide

In a 50-mL round-bottom flask was combined the compound from theprevious step (150 mg, 0.23 mmol, 1.00 equiv), CF₃COOH (1 mL), andCH₂Cl₂ (10 mL). The solution was stirred for 12 h at room temperature,then concentrated under reduced pressure and purified by Prep-HPLC,affording 63.1 mg (42%) of 12 trifluoroacetic acid salt as a whitesolid.

Example 20

Synthesis of 20

Ethyl 4-methanesulfonylphenyl benzoate

In a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined a solution of ethyl 4-bromobenzoate(5 g, 21.83 mmol, 1.00 equiv) in dioxane (200 mL),(4-methanesulfonylphenyl)boronic acid (5.2 g, 26.00 mmol, 1.19 equiv),and a solution of K₂CO₃ (6 g, 43.41 mmol, 1.99 equiv) in water (20 mL),Pd(PPh₃)₄ (2.5 g). The resulting solution was stirred for 16 h at 100°C. in an oil bath, then cooled to room temperature, diluted with 500 mLof H₂O, and extracted with 3×200 mL of EtOAc. The organic layers werecombined, washed with 1×500 mL of brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified byre-crystallization from EtOAc to afford 6 g of the product as a whitesolid.

4-Methanesulfonylphenyl benzoic acid (209)

In a 250-mL round-bottom flask were combined the product from theprevious reaction (9 g, 29.57 mmol, 1.00 equiv), methanol (150 mL), NaOH(3 g, 75.00 mmol, 2.54 equiv). The resulting solution was stirred for 5h at 25° C., then concentrated under vacuum and diluted with 200 mL ofH₂O. The pH value of the aqueous was adjusted to 2 with HCl (2 M). Thesolids that formed were collected by filtration and dried in an oven,affording 6 g (73%) of the product as a white solid.

4-Methanesulfonylphenyl benzoyl chloride

The method used to prepare 201 was used with the compound from theprevious step (5 g, 18.10 mmol), affording 5 g (94%) of the product as alight yellow solid.

N—((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4-(methylsulfonyl)piperazin-1-yl)-1-oxobutan-2-yl)-4′-(methylsulfonyl)biphenyl-4-carboxamide

The remainder of the synthesis proceeded as for Scheme 2.

Example 21

Synthesis of 21

4-(Methoxycarbonyl)benzoyl chloride

The method used to prepare 201 was used with 4-(methoxycarbonyl)benzoicacid (2 g, 11.10 mmol,) to afford 2.2 g (100%) of the product as ayellow solid.

N-(2,2-Diethoxyethyl)-4-(methoxycarbonyl)benzamide

The method used to prepare 203 was used with the compound from theprevious step and 2,2-diethoxyethan-1-amine (1.22 g, 9.16 mmol) toafford 3 g (92%) of the product as a yellow solid.

Methyl 4-(oxazol-2-yl)-benzoate

Into a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed the product from the previous step (3g, 10.16 mmol, 1.00 equiv), methanesulfonic acid (150 mL), phosphoruspentoxide (8.52 g, 59.18 mmol, 5.00 equiv). The resulting solution wasstirred for 2 h at 140° C. in an oil bath, cooled to room temperature,and extracted with 3×150 mL of EtOAc The organic layers were combined,dried over Na₂SO₄, concentrated under reduced pressure, and applied ontoa silica gel column with EtOAc/petroleum ether (1:1), affording 1.5 g(73%) of the product as a yellow solid.

4-(Oxazol-2-yl)-benzoic acid

The method used to prepare 209 was used with the product from theprevious step (1.5 g, 7.38 mmol, 1.00 equiv) to afford 1.2 g (86%) ofthe product as a yellow solid.

N—((S)-4-((1R,2S)-2-(4-Fluorophenyl)cyclopropylamino)-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl)-4-(oxazol-2-yl)phenyl-4-carboxamide

The remainder of the synthesis proceeded as for Scheme 2.

Example 23

Synthesis of 23

(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](propen-3-yl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]pentanoicacid

The method used to prepare 203 was used with(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-aminopentanoicacid (800 mg, 2.61 mmol, 1.00 equiv) and4-(1H-1,2,3-triazol-1-yl)benzoyl chloride (800 mg, 3.85 mmol, 1.50equiv) to afford 550 mg (44%) of the product as a yellow solid.

N-[(2S)-1-(4-(tert-butyloxycarbonyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 206 was used with the compound from theprevious step (250 mg, 0.52 mmol, 1.00 equiv) and4-(tert-butyloxycarbonyl)piperazine (148 mg, 0.79 mmol, 1.50 equiv) toafford 210 mg (62%) of the product as a yellow solid.

N-[(2S)-1-(4-(tert-butyloxycarbonyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(210)

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was the compound from the previous step (210 mg,0.32 mmol, 1.00 equiv), THF (30 mL), barbituric acid (127 mg, 0.81 mmol,2.50 equiv), and tetrakis(triphenylphosphine)-palladium (94 mg, 0.08mmol, 0.25 equiv). The resulting solution was stirred for 2 h at 50° C.in an oil bath, then concentrated under reduced pressure, applied onto asilica gel column with CH₂Cl₂/methanol (10:1), affording 150 mg (76%) ofthe product as a yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-(1H-1,2,3-triazol-1-yl)pentan-2-yl]-4-(pyrimidin-2-yl)benzamidetrifluoroacetic acid salt

The procedure used to prepare 1 was used with the compound produced inthe previous step (150 mg, 0.25 mmol, 1.00 equiv) to afford 53.2 mg(43%) of the product as an off-white solid.

Example 35

Synthesis of 35

N-[(2S)-1-(3-cyanoazetidin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-oxopentan-2-yl]-4-(1H-pyrazol-1-yl)benzamide

The method used to prepare 206 was used with(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(1H-pyrazol-1-yl)phenyl]-formamido]pentanoicacid (400 mg, 0.84 mmol, 1.00 equiv) and azetidine-3-carbonitrile toafford 301 mg (66%) of the product as a light yellow oil.

N-[(2S)-1-(3-cyanoazetidin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-pyrazol-1-yl)benzamide(35)

The method used to prepare 210 was used with the compound produced inthe previous step (301 mg, 0.56 mmol, 1.00 equiv) to afford 72.1 mg(26%) of 35 as an off-white solid.

Example 86

Synthesis of 86

N-[(2S)-1-(4-(tert-butyloxycarbonyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide

The method used to prepare 206 was used with(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanoicacid (250 mg, 0.51 mmol, 1.00 equiv) and1-tert-butyl-1^3,3,6-oxadiazocan-2-one (143 mg, 0.76 mmol, 1.50 equiv)to afford 280 mg (66%) of the product as a yellow solid.

N-[(2S)-1-(4-(tert-butyloxycarbonyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide

The method used to prepare 210 was used with the compound produced inthe previous step (280 mg, 0.43 mmol) to afford 0.2 g (76%) of theproduct as a yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxo-1-(piperazin-1-yl)pentan-2-yl]-4-(pyrimidin-2-yl)benzamidetrifluoroacetic acid salt (86)

The method used to prepare 1 was used with the compound produced in theprevious step (200 mg, 0.32 mmol) to afford 70 mg (34%) of 86 as a whitesolid.

Example 91

Synthesis of 91

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-carboethoxypiperidine-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 206 was used with(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]-formamido]pentanoicacid (200 mg, 0.42 mmol, 1.00 equiv) and methyl piperidine-4-carboxylate(80 mg, 0.56 mmol, 1.33 equiv) to afford 180 mg (71%) of the product asa colorless oil.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-carboethoxypiperidine-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 210 was used with the compound produced inthe previous step (180 mg, 0.30 mmol) to afford 100 mg (60%) of theproduct as an orange oil.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-carboxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(91)

In a 50-mL round-bottom flask were combined the compound from theprevious step (100 mg, 0.17 mmol, 1.00 equiv) in THF (20 mL) and LiOH(20 mg, 0.84 mmol, 4.82 equiv) in water (5 mL). The resulting solutionwas stirred for 16 h at 25° C., then concentrated under vacuum. Thecrude product was purified by Prep-HPLC, affording 52.9 mg (46%) of theproduct as a light yellow solid.

Example 92

Synthesis of 92

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]benzamide(211)

In a 100-mL round-bottom flask were combined a solution of4-cyanobenzoic acid (136.7 mg, 0.93 mmol, 1.20 equiv) CH₂Cl₂ (20 mL),i-Pr₂NEt (298 mg, 2.31 mmol, 3.00 equiv), and1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (“HATU”) (439 mg, 1.15 mmol, 1.50 equiv),The mixture was stirred for 1 h, then a solution of(2S)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)-amino]-1-(4-hydroxypiperidin-1-yl)pentan-1-one(300 mg, 0.77 mmol, 1.00 equiv) in CH₂Cl₂ (5 mL) was added. Theresulting solution was stirred for 60 min at 25° C., then concentratedunder vacuum and applied onto a silica gel column with CH₂Cl₂/methanol(10:1), affording 480 mg (120%) of the product as a light yellow liquid.

4-Cyano-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]benzamide(92)

The method used to prepare 210 was used with the compound produced inthe previous step (480 mg, 0.93 mmol) to afford 46.7 mg (11%) of 92 as alight yellow solid.

Example 109

Synthesis of 109

Methyl4-[[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]carbamoyl]benzoate

The method used to prepare 211 was used with 4-(methoxycarbonyl)benzoicacid (231.4 mg, 1.28 mmol, 1.00 equiv) and(2S)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)pentan-1-one(500 mg, 1.28 mmol, 1.00 equiv) to afford 420 mg (59%) of the product asa light yellow oil.

Methyl4-[[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]carbamoyl]benzoate(109)

The method used to prepare 210 was used with the compound produced inthe previous step (420 mg, 0.76 mmol) to afford 650 mg (99%) of theproduct as an orange oil.

4-[[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]carbamoyl]benzoicacid (109)

In a 100-mL round-bottom flask were combined the compound from theprevious step (650 mg, 1.27 mmol, 1.00 equiv), LiOH (60 mg, 2.51 mmol,2.00 equiv), methanol (20 mL), and water (13 mL). The resulting solutionwas stirred for 3 h at room temperature. The resulting mixture wasconcentrated under vacuum. The crude product was purified by Prep-HPLCto afford 45.1 mg (7%) of the product as an orange solid.

Example 109

Synthesis of 110

N-[(2R)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]carbamate

The method used to prepare 211 was used with2-[[(tert-butoxy)carbonyl]amino]-5-[[2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid (1 g, 2.46 mmol, 1.00 equiv) and 1-methylpiperazine (500 mg, 4.99mmol, 2.00 equiv) to afford in 1.1 g (95%) of the product as off-whiteoil.

4-(1H-1,2,3-Triazol-1-yl)benzoic acid (926 mg, 4.90 mmol)(2R)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)pentan-1-one

The procedure used to prepare 1 was used with the compound produced inthe previous step (2.4 g, 4.91 mmol) to afford 1.8 g (95%) of theproduct as off-white oil.

N-[(2R)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 211 was used with the compound produced inthe previous step (1.9 g, 4.89 mmol, 1.00 equiv) to afford 1.2 g (44%)of the product as off-white oil.

N-[(2R)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(110)

The method used to prepare 210 was used with the compound produced inthe previous step (1.2 g, 2.14 mmol) to afford 15.1 mg (1%) of 110 as awhite solid.

Example 111

Synthesis of 111

(2S)-2-[[(tert-Butoxy)carbonyl]amino]-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid

In a 250-mL round-bottom flask were combined(2S)-2-amino-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid; CF₃COOH (2 g, 4.76 mmol, 1.00 equiv), 1,4-dioxane (100 mL), Boc₂O(1.44 g, 6.60 mmol, 1.39 equiv), and a solution of Na₂CO₃ (1.4 g, 13.21mmol, 2.78 equiv) in H₂O (25 mL). The resulting solution was stirred for1 h at room temperature, concentrated under vacuum, diluted with 40 mLof DMF and applied onto a C18 column with MeCN/H₂O (1:1), affording 1.8g (93%) of the product as light yellow oil.

tert-ButylN-[(2S)-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]carbamate

The method used to prepare 206 was used with(2S)-2-[[(tert-butoxy)carbonyl]amino]-5-[[(2S)-2-(4-fluorophenyl)-cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid (900 mg, 2.21 mmol, 1.00 equiv) and piperidin-4-ol (335 mg, 3.31mmol, 1.50 equiv) to afford 1.2 g (98%) of the product as light yellowoil.

(2S)-2-Amino-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)pentan-1-one;trifluoroacetic acid salt

The method used to prepare 1 was used with the compound produced in theprevious step (1.2 g, 2.45 mmol, 1.00 equiv) to afford 880 mg (71%) ofthe product as a yellow oil.

N-[(2S)-5-[[(2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 211 was used with the compound produced inthe previous step (400 g, 794.39 mmol, 1.00 equiv), and4-(1H-1,2,3-triazol-1-yl)benzoic acid (205 mg, 1.08 mmol) to afford 300mg of the product as a yellow solid.

N-[(2S)-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-hydroxypiperidin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(111)

The method used to prepare 210 was used with the compound produced inthe previous step (300 mg, 0.54 mmol) to afford (23%) of 111 as anoff-white solid.

Example 117

Synthesis of 117

tert-ButylN-[(2R)-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-oxo-1-(3-(dimethylamino)-azetidin-1-yl)-pentan-2-yl]carbamate

The method used to prepare 206 was used with(2S)-2-[[(tert-butoxy)carbonyl]amino]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid (400 mg, 0.98 mmol, 1.00 equiv) and N,N-dimethylazetidin-3-amine(147.9 mg, 1.48 mmol, 1.50 equiv) to afford 490 mg (99%) of the productas an off-white solid.

1-Oxo-1-(3-(dimethylamino)-azetidin-1-yl)-(2R)-2-amino-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-pentane

The procedure used to prepare 1 was used with the compound produced inthe previous step (490 mg, 1.00 mmol, 1.00 equiv) to afford 470 mg (99%)of the product as a light yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(3-(dimethylamino)-azetidin-1-yl-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)-benzamide

The method used to prepare 211 was used with the compound produced inthe previous step (470 mg, 1.21 mmol, 1.00 equiv). to afford 608 mg(90%) of the product as a light yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(3-(dimethylamino)-azetidin-1-yl-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)-benzamide(117)

The method used to prepare 210 was used with the compound produced inthe previous step (978 mg, 1.75 mmol, 1.00 equiv) to afford 61.0 mg (7%)of 117 as an off-white solid.

Example 126

Synthesis of 126

2-[4-[(2S)-2-[(4-Cyanophenyl)formamido]-5-[[(1R,2S)-2-(4-fluorophenyl)-cyclopropyl]amino]pentanoyl]piperazin-1-yl]acetate(126)

In a 100-mL round-bottom flask were combined a solution of ethyl2-[4-[(2S)-2-[(4-cyanophenyl)formamido]-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]pentanoyl]piperazin-1-yl]acetate(150 mg, 0.27 mmol, 1.00 equiv) in THF (20 mL) and a solution of LiOH(13.1 mg, 0.55 mmol, 2.00 equiv) in water (5 mL). The resulting solutionwas stirred for 2 h at 25° C. The pH value of the solution was adjustedto 3-4 with aqueous HCl (2 M). The resulting mixture was concentratedunder vacuum. The residue was applied onto a reverse-phase silica gelcolumn with MeCN/H₂O (1:10) The crude product (5 mL) was purified byPrep-HPLC to afford 108 mg (64%) of 126 as a light yellow solid.

Example 127

Synthesis of 127

N-BenzylN-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]-N-[(4R)-5-(4-methylpiperazin-1-yl)-5-oxo-4-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]pentyl]carbamate(127)

In a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was combined a solution ofN-[(2R)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(40 mg, 0.08 mmol, 1.00 equiv) in tetrahydrofuran (5 mL), TEA (23 mg,0.23 mmol, 3.00 equiv), CbzCl (19.7 mg, 0.12 mmol, 1.50 equiv). Theresulting solution was stirred for 1 h at room temperature, and was thenquenched by the addition of 15 mL of water/ice, extracted with 3×20 mLof EtOAc. The organic layers were combined, washed with 3×20 mL ofbrine, dried over Na₂SO₄ and concentrated under vacuum. The residue wasapplied onto a silica gel column with dichloromethane/methanol (20:1) toafford 49.5 mg (98%) of 127 as an off-white solid.

Example 128

Synthesis of 128

(2S)-5-Methoxy-5-oxo-2-[(4-phenylphenyl)formamido]pentanoic acid

The method used to prepare 203 was used with(2S)-2-amino-5-methoxy-5-oxopentanoic acid (4.9 g, 30.41 mmol, 1.00equiv) and 4-phenylbenzoyl chloride (5 g, 23.08 mmol, 0.76 equiv) toafford 4 g (39%) of the product as a white solid.

Methyl(4S)-5-(4-methanesulfonylpiperazin-1-yl)-5-oxo-4-[(4-phenylphenyl)formamido]pentanoate

The method used to prepare 206 was used with(2S)-5-methoxy-5-oxo-2-[(4-phenylphenyl)formamido]pentanoic acid (2 g,5.86 mmol, 1.00 equiv) and 1-methanesulfonylpiperazin (1.15 g, 7.00mmol, 1.20 equiv) to afford 2.5 g (88%) of the product as an off-whitesolid

(4S)-5-(4-Methanesulfonylpiperazin-1-yl)-5-oxo-4-[(4-phenylphenyl)formamido]pentanoicacid

Into a 250-mL round-bottom flask, were combined the compound from theprevious step (1 g, 2.05 mmol, 1.00 equiv), LiOH (73 mg, 3.05 mmol, 1.49equiv), and THF/H₂O (50/12 mL). The resulting solution was stirred for 1h at room temperature. The pH value of the solution was adjusted to 2with HCl (2 M). The resulting solution was extracted with 3×30 mL ofEtOAc, and the organic layers were combined and dried over Na₂SO₄ andconcentrated under vacuum, affording 700 mg (72%) of the product as anoff-white solid.

(4S)-5-(4-Methanesulfonylpiperazin-1-yl)-5-oxo-4-[(4-phenylphenyl)formamido]-1-pentanol(202)

Into a 100-mL round-bottom flask were combined a solution of thecompound from the previous step (700 mg, 1.48 mmol, 1.00 equiv) in THF(10 mL) and BH₃ (1 M in THF) (2.2 mL). The resulting solution wasstirred for 2 h at room temperature. The reaction was then quenched bythe addition of 20 mL of water, then extracted with 3×20 mL of EtOAc.The organic layers were combined, dried over Na₂SO₄, and concentratedunder vacuum. The residue was applied onto a silica gel column withEtOAc/petroleum ether (1:1) to afford 400 mg (59%) of the product as ayellow oil.

(4S)-5-(4-Methanesulfonylpiperazin-1-yl)-5-oxo-4-[(4-phenylphenyl)formamido]pentylmethanesulfonate

In a 50-mL round-bottom flask were combined a solution of the compoundfrom the previous step (300 mg, 0.65 mmol, 1.00 equiv) and Et₃N (132 mg,1.30 mmol, 2.00 equiv) in CH₂Cl₂ (5 mL). This was followed by theaddition of methanesulfonyl chloride (90 mg, 0.78 mmol, 1.20 equiv)dropwise with stirring at 0° C. The resulting solution was stirred for30 min at room temperature. The reaction was then quenched by theaddition of 10 mL of water. The resulting solution was extracted with3×10 mL of CH₂Cl₂, and the organic layers were combined and dried overanhydrous Na₂SO₄ and concentrated under vacuum. The residue was appliedonto a silica gel column with EtOAc/petroleum ether (1:2) to afford 200mg (57%) of the product as an off-white solid.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methanesulfonylpiperazin-1-yl)-1-oxopentan-2-yl]-4-phenylbenzamide(128)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined the compound from the previous step(200 mg, 0.37 mmol, 1.00 equiv), iPr₂NEt (96 mg, 0.74 mmol, 2.00 equiv),KI (62 mg), (1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (62 mg, 0.41mmol, 1.10 equiv) and CH₃CN (10 mL). The resulting solution was stirredfor 12 h at 50° C. in an oil bath. The reaction was then quenched by theaddition of 10 mL of water. The resulting solution was extracted with3×10 mL of EtOAc, and the organic layers were combined and dried overanhydrous Na₂SO₄ and concentrated under vacuum. The crude product waspurified by Prep-HPLC, affording 12.7 mg (6%) of 128 as an off-whitesolid.

Example 129

Synthesis of 129

(2S)-5-Methoxy-5-oxo-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]]-pentanoicacid

The method used to prepare 203 was used with(2S)-2-amino-5-methoxy-5-oxopentanoic acid (3 g, 18.63 mmol, 1.00equiv), and 4-(1H-1,2,3-triazol-1-yl)benzoyl chloride (4.65 g, 22.36mmol, 1.20 equiv) to afford 2.5 g (40%) of the product as a yellow oil.

Methyl(4S)-5-(4-methylpiperazin-1-yl)-5-oxo-4-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido-]-pentanoate

The method used to prepare 206 was used with the compound produced inthe previous step (2.5 g, 7.53 mmol, 1.00 equiv) in and1-methylpiperazine (1.13 g, 11.30 mmol, 1.50 equiv) to afford 2 g (64%)of the product as a yellow oil.

(4S)-5-(4-Methylpiperazin-1-yl)-5-oxo-4-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]-pentanoicacid

The method to prepare 205 was used with the compound produced in theprevious step (2 g, 4.83 mmol) to afford 1.8 g (93%) of the product asan off-white solid.

(4S)-5-(4-Methylpiperazin-1-yl)-5-oxo-4-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]-1-pentanol

In a 100-mL round-bottom flask were combined a solution of the productfrom the previous step (1.6 g, 4.00 mmol, 1.00 equiv) in THF (20 mL),N-methyl morpholine (983 mg, 8.40 mmol, 2.10 equiv), and tert-butylchloroformate (1151 mg, 8.40 mmol, 2.10 equiv). The resulting mixturewas stirred for 2 h at −20 degrees, then NaBH₄ (1.52 g, 40.0 mmol, 10.00equiv) in methanol (10 mL) was added. The resulting solution was stirredfor 2 h at −20° C. in a liquid nitrogen bath. The resulting mixture wasconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (10:1), affording 450 mg (29%)of the product as an off-white solid.

(4S)-5-(4-Methylpiperazin-1-yl)-5-oxo-4-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]-pentanal

The method used to prepare 208 was used with the compound produced inthe previous step (200 mg, 0.52 mmol, 1.00 equiv) to afford 150 mg (75%)of the product as an off-white solid.

N-[(2S)-5-[[(1R,2S)-2-(4-methoxyphenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(129)

In a 25-mL round-bottom flask were combined solution of the compoundfrom the previous step (150 mg, 0.391 mmol, 1.00 equiv),(1R,2S)-2-(4-methoxyphenyl)cyclopropanamine (76 mg, 0.47 mmol, 1.20equiv), NaBH(OAc)₃ (199 mg, 0.94 mmol, 2.40 equiv) and CH₂Cl₂ (20 mL).The resulting solution was stirred for 10 min at 25° C., then dilutedwith 30 mL of H₂O and extracted with 2×20 mL of CH₂Cl₂. The organiclayers were combined, dried over Na₂SO₄, and concentrated under vacuum.The crude product was purified by Prep-HPLC, affording 5.9 mg (2.8%) of129 as an off-white solid.

Example 130

Synthesis of 130

Methyl 4-azido-2-fluorobenzoate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of methyl4-amino-2-fluorobenzoate (1 g, 5.91 mmol, 1.00 equiv) in hydrogenchloride (5 mL), a solution of NaNO₂ (407 mg, 5.90 mmol, 1.00 equiv) inwater (5 mL), a solution of NaN₃ (575 mg, 8.85 mmol, 1.50 equiv) inwater (5 mL). The resulting solution was stirred for 15 min at 0° C. Thesolids were filtered out. The resulting mixture was concentrated undervacuum. The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:1). This resulted in 800 mg (69%) of theproduct as a yellow solid.

Methyl 2-fluoro-4-[5-(trimethylsilyl)-1H-1,2,3-triazol-1-yl]benzoate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of methyl4-azido-2-fluorobenzoate (800 mg, 4.10 mmol, 1.00 equiv) in methanol (20mL), ethynyltrimethylsilane (603 mg, 6.14 mmol, 1.50 equiv), CuI (1.2 g,6.30 mmol, 1.50 equiv), TEA (1.2 g, 11.88 mmol, 3.00 equiv). Theresulting solution was stirred for 16 h at room temperature. Thereaction was then quenched by the addition of 50 mL of water/ice. Theresulting solution was extracted with 3×50 mL of ethyl acetate and theorganic layers combined and concentrated under vacuum. The residue wasapplied onto a silica gel column with ethyl acetate/petroleum ether(1:1). This resulted in 700 mg (58%) of the product as a yellow solid.

Methyl 2-fluoro-4-(1H-1,2,3-triazol-1-yl)benzoate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of methyl2-fluoro-4-[5-(trimethylsilyl)-1H-1,2,3-triazol-1-yl]benzoate (700 mg,2.39 mmol, 1.00 equiv) in tetrahydrofuran (10 mL), TBAF (1.25 g, 4.78mmol, 2.00 equiv), AcOH (144 mg, 2.40 mmol, 1.00 equiv). The resultingsolution was stirred for 2 days at room temperature. The reaction wasthen quenched by the addition of 50 mL of water/ice. The resultingsolution was extracted with 3×50 mL of ethyl acetate and the organiclayers combined and concentrated under vacuum. The residue was appliedonto a silica gel column with dichloromethane/methanol (100:1). Thisresulted in 550 mg (crude) of the product as a yellow solid.

2-Fluoro-4-(1H-1,2,3-triazol-1-yl)benzoic acid

The procedure used to prepare 205 was used with the compound produced inthe previous step to afford 450 mg (87%) of the product as a lightyellow solid

2-Fluoro-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 211 was used with the compound produced inthe previous step (160 mg, 0.77 mmol, 1.00 equiv) and(2S)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)pentan-1-one(300 mg, 0.77 mmol, 1.00 equiv) to afford 250 mg (56%) of the product asa yellow solid.

2-Fluoro-N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(130)

The method used to prepare 210 was used with the compound produced inthe previous step (250 mg, 0.43 mmol) to afford 33.2 mg (14%) of 130 asa light brown solid.

Example 131

Synthesis of 131

tert-Butyl 4-carbamoyl-4-fluoropiperidine-1-carboxylate

In a 250-mL round-bottom flask were combined1-[(tert-butoxy)carbonyl]-4-fluoropiperidine-4-carboxylic acid (3 g,12.13 mmol, 1.00 equiv), DMF (50 mL), NH4Cl (1.75 g, 32.72 mmol, 1.50equiv), HATU (9.23 g, 24.27 mmol, 2.00 equiv), and iPr₂NEt (3.13 g,24.22 mmol, 2.00 equiv). The resulting solution was stirred overnight at25° C. The resulting solution was extracted with 200 mL of EtOAc and theorganic layers were combined, washed with 5×50 mL of H₂O, then appliedonto a silica gel column with ethyl acetate/petroleum ether, affording2.7 g (90%) of the product as a white solid.

tert-butyl 4-cyano-4-fluoropiperidine-1-carboxylate

In a 250-mL round-bottom flask were combined the compound from theprevious step (1.6 g, 6.50 mmol, 1.00 equiv), pyridine (15 mL), TFAA (10mL), and THF (30 mL). The resulting solution was stirred for 1 h at 0°C. in a water/ice bath. The crude product (50 mL) was purified byFlash-Prep-HPLC with the following conditions (IntelFlash-1): Column,C18 silica gel; mobile phase, MeCN/H₂O=0/100 increasing toMeCN/H₂O=50/50 within 30 min; Detector, UV 254 nm. 10 mL product wasobtained, affording 1.2 g (81%) of the product as a white solid.

4-Fluoropiperidine-4-carbonitrile

In a 250-mL round-bottom flask were combined the compound from theprevious step (800 mg, 3.50 mmol, 1.00 equiv), CF₃COOH (1 mL), andCH₂Cl₂ (5 mL). The resulting solution was stirred for 1 h at 25° C.,then concentrated under vacuum to afford 350 mg (78%) of the product asa white solid.

N-[(2S)-1-(4-Cyano-4-fluoropiperidin-1-yl)-1-oxo-5-[[(1R,2S)-2-phenylcyclopropyl](prop-2-en-1-yl)amino]pentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 206 was used with(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]pentanoicacid (700 mg, 1.47 mmol, 1.00 equiv) and4-fluoropiperidine-4-carbonitrile (350 mg, 2.73 mmol, 2.00 equiv) toafford 816 mg (98%) of the product as a yellow oil.

N-[(2S)-1-(4-Cyano-4-fluoropiperidin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(131)

The method used to prepare 210 was used with the compound from theprevious step to afford 40.7 mg (5%) of the product as a white solid.

Example 132

Synthesis of 132

(4S)—N-Methyl-N-methoxy-5-(tert-butoxy)-4-[(tert-butoxycarbonyl)amino]-5-oxopentanamide

In a 1-L round-bottom flask were combined a solution of(4S)-5-(tert-butoxy)-4-[[(tert-butoxy)carbonyl]amino]-5-oxopentanoicacid (20.0 g, 65.93 mmol, 1.00 equiv) in CH₂Cl₂ (200 mL), EDCI (18.9 g,98.59 mmol, 1.50 equiv), HOBT (13.4 g, 98.80 mmol, 1.50 equiv),methoxy(methyl)amine hydrochloride (9.7 g, 98.94 mmol, 1.50 equiv), andEt₃N (20.0 g, 197.65 mmol, 3.00 equiv). The resulting solution wasstirred for 3 h at 25° C. The then washed with 3×100 mL of H₂O, driedover Na₂SO₄, concentrated under vacuum, then applied onto a silica gelcolumn with EtOAc/petroleum ether (1:3) to afford 22.1 g (96%) of theproduct as a light yellow oil.

(4S)—N-Methyl-N-methoxy-5-(tert-butoxy)-4-[di(tert-butoxycarbonyl)amino]-5-oxopentanamide

The method used to prepare 204 was used with the compound from theprevious step (19.1 g, 54.99 mmol, 1.00 equiv) to afford 23.6 g (96%) ofthe product as a light yellow oil.

(4S)-5-(tert-Butyloxy)-4-[di(tert-butoxycarbonyl)amino]-5-oxopentanal

Into a 500-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen were combined a solution of the compoundfrom the previous step (6 g, 13.42 mmol, 1.00 equiv) in THF (50 mL),followed by the addition of diisobutyl aluminium hydride (30 mL, 2.50equiv) dropwise with stirring at −78° C. in 30 min. The resultingsolution was stirred for 30 min at −78° C., then quenched by theaddition of 50 mL NH₄Cl(aq). The resulting solution was extracted with3×100 mL of EtOAc. The organic layers were combined, dried over Na₂SO₄and concentrated under vacuum, affording 6.3 g (crude) of the product asa light yellow oil.

tert-Butyl(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-2-[di(tert-butoxycarbonyl)amino]pentanoate

Into a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined the compound from the previous step(6.3 g, 16.26 mmol, 1.00 equiv), 2-(4-fluorophenyl)cyclopropan-1-aminehydrochloride (2.45 g, 13.06 mmol, 0.80 equiv), NaBH(OAc)₃ (8.25 g,38.93 mmol, 2.40 equiv), and methanol (150 mL). The resulting solutionwas stirred for 30 min at 25° C. The resulting mixture was concentratedunder vacuum, then diluted with 100 mL EtOAc. The resulting mixture waswashed with 3×20 mL of H₂O, dried and concentrated under vacuum. Theresidue was applied onto a silica gel column with EtOAc/petroleum ether(1:4), affording 3.1 g (36%) of the product as a light yellow oil.

tert-Butyl(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](propen-2-yl)amino]-2-[di(tert-butoxycarbonyl)amino]pentanoate

Into a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined a solution of the compound from theprevious step (3.1 g, 5.93 mmol, 1.00 equiv) in DMF (10 mL), K₂CO₃ (2.45g, 17.78 mmol, 3.00 equiv), and 3-bromoprop-1-ene (1.43 g, 11.85 mmol,2.00 equiv). The resulting solution was stirred for 2 h at 25° C. Theresulting solution was diluted with 50 mL of EtOAc, then washed with 3×5mL of H₂O, dried over Na₂SO₄, and concentrated under vacuum. The residuewas applied onto a silica gel column with EtOAc/petroleum ether (1:7) toafford 3 g (90%) of the product as a light yellow oil.

(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](propen-2-yl)amino]-2-aminopentanoicacid

In a 500-mL round-bottom flask were combined the compound from theprevious step (3 g, 43.01 mmol, 1.00 equiv) and CF₃COOH (10 mL). Theresulting solution was stirred for 16 h at 25° C. then concentratedunder vacuum. The crude product was purified by Flash-Prep-HPLC withCH₃CN:H₂O (1:100-15:1) to afford 2 g (83%) of the product as a lightyellow solid.

(2S)-5-[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](propen-2-yl)amino]-2-[(tert-butoxycarbonyl)amino]pentanoicacid

Into a 500-mL round-bottom flask, was placed a solution of the compoundfrom the previous step (2 g, 6.51 mmol, 1.00 equiv) in dioxane (20 mL),(Boc)₂O (2.13 g, 9.77 mmol, 1.50 equiv), Na₂CO₃ (2.07 g, 19.54 mmol, 3.0equiv) in H₂O (20 ml). The resulting solution was stirred for 2 h atroom temperature. The solid was removed by filtration, and the resultingsolution was concentrated under vacuum. The crude product was purifiedby flash with CH₃CN:H₂O (1:100-1:1) to afford 2.3 g (86%) of the productas a yellow oil.

(2S)-2-[(tert-butoxycarbonyl)amino]-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](propen-2-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentane

The method used to prepare 206 was used with the compound from theprevious step (2.3 g, 5.67 mmol, 1.00 equiv) and N-methylpiperazine (850mg, 8.50 mmol, 1.5 equiv) to afford 2.3 g (83%) of PH-IMA-2013-003-384-8as a yellow oil.

(2S)-2-[(tert-butoxycarbonyl)amino]-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](propen-2-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentane

The method used to prepare 211 was used with the compound from theprevious step to afford 1.8 g of the product as a yellow oil.

(2S)—N-[5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](propen-2-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 211 was used with the compound from theprevious step (1.8 g, 4.63 mmol, 1.0 equiv) and4-(1H-1,2,3-triazol-1-yl)benzoic acid (875 mg, 4.63 mmol, 1.0 equiv) toafford 1.7 g (66%) of the product as a yellow solid.

(2S)—N-[5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(132)

The method used to prepare 210 was used with the compound from theprevious step to afford 281.8 mg (18%) of 132 as a light-yellow solid.

Example 134

Synthesis of 134

Methyl 4-(2-pyrazinyl) benzoate

In a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined a solution of[4-(methoxycarbonyl)phenyl]boronic acid (1 g, 5.56 mmol, 1.20 equiv) in1,4-dioxane (30 mL), 2-bromopyrazine (800 mg, 5.03 mmol, 1.00 equiv), asolution of Na₂CO₃ (1.5 g, 14.15 mmol, 3.00 equiv) in water (30 mL), andPd(Ph₃P)₂Cl₂ (330 mg, 0.47 mmol, 0.10 equiv). The resulting solution wasstirred for 16 h at 90° C. in an oil bath. The solids were filtered out.The resulting solution was extracted with 3×30 mL of EtOAc, and theorganic layers combined and concentrated under vacuum. This resulted in0.8 g (74%) of the product as a white solid.

4-(2-Pyrazinyl) benzoic acid

In a 100-mL round-bottom flask were combined a solution of methyl4-(pyrazin-2-yl)benzoate (800 mg, 3.73 mmol, 1.00 equiv) in methanol (20mL), a solution of NaOH (150 mg, 3.75 mmol, 1.00 equiv) in water (20mL). The resulting solution was stirred for 16 h at 80° C. in an oilbath. The pH value of the solution was adjusted to 7 with HCl (1 M). Thesolids were collected by filtration. The resulting mixture wasconcentrated under vacuum. This resulted in 0.65 g (87%) of the productas a white solid.

4-(2-Pyrazinyl)-benzoyl chloride

In a 100-mL round-bottom flask were combined 4-(pyrazin-2-yl)benzoicacid (650 mg, 3.25 mmol, 1.00 equiv) and thionyl chloride (20 mL). Theresulting solution was stirred for 16 h at 80° C. in an oil bath. Theresulting mixture was concentrated under vacuum. This resulted in 0.7 g(99%) of the product as a light yellow solid.

(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(2-pyrazinyl)phenyl]formamido]pentanoicacid

The method used to prepare 203 was used with the compound produced inthe previous step (500 mg, 1.63 mmol, 1.00 equiv) and4-(pyridin-2-yl)benzoyl chloride (393 mg, 1.81 mmol, 1.10 equiv), toafford 360 mg (45%) of the product as a yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrazin-2-yl)benzamide

The method used to prepare 206 was used with the compound produced inthe previous step (360 mg, 0.74 mmol, 1.00 equiv) and 1-methylpiperazine(111 mg, 1.11 mmol, 1.50 equiv) to afford 300 mg (71%) of the product asa yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrazin-2-yl)benzamide(134)

The method used to prepare 210 was used with the compound produced inthe previous step (300 mg, 0.53 mmol, 1.00 equiv), affording 88.8 mg(32%) of the product as an off-white solid.

Example 135

Synthesis of 135

Methyl 4-(pyrimidin-5-yl)benzoate

In a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined solution of 5-bromopyrimidine (2.2g, 13.84 mmol, 1.10 equiv) in MeCN (60 mL). This was followed by theaddition of [4-(methoxycarbonyl)phenyl]boronic acid (2.26 g, 12.56 mmol,1.00 equiv), in portions. To this was added a solution of Na₂CO₃ (2.9 g,27.36 mmol, 2.00 equiv) in water (30 mL) dropwise with stirring at roomtemperature in 2 min. To the mixture was added Pd(PPh₃)₄ (1.45 g, 1.25mmol, 0.10 equiv), in portions. The resulting solution was stirred for 4h at 90° C. The solids were removed by filtration. The resulting mixturewas concentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:1). This resulted in 1.31 g(49%) of the product as a yellow solid.

4-(Pyrimidin-5-yl)benzoic acid

The method used to prepare 209 was used with the compound produced inthe previous step (1.31 g, 6.12 mmol, 1.00 equiv) to afford 0.7 g (57%)of as a white solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrimidin-5-yl)benzamide

The method used to prepare 211 was used with the compound produced inthe previous step (250 mg, 0.50 mmol, 1.00 equiv) to afford (70%) of asa yellow foam.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrimidin-5-yl)benzamide(135)

The method used to prepare 210 was used with the compound produced inthe previous step (200 mg, 0.35 mmol,) to afford 15 mg (8%) of 135 as anoff-white solid.

Example 137

Synthesis of 137

tert-Butyl 4-(d₃)-methylpiperazine-1-carboxylate

In a 100-mL round-bottom flask were combined tert-butylpiperazine-1-carboxylate (1 g, 5.37 mmol, 1.00 equiv), K₂CO₃ (2.23 g,16.13 mmol, 3.01 equiv) and THF (40 mL). The resulting solution wasstirred for 1 h at room temperature. This was followed by the additionof iodomethane-d₃ (780 mg, 5.38 mmol, 1.00 equiv) dropwise with stirringat −12° C. The resulting solution was stirred overnight at roomtemperature. The solids were removed by filtration, and the crudeproduct was purified by Prep-HPLC. The resulting solution was extractedwith 3×30 mL of 5:1 CH₂Cl₂:MeOH, and the organic layers were combinedand concentrated under vacuum, affording 500 mg (46%) of the product asa colorless oil.

1-(d₃)-Methylpiperazine

The method used to prepare 1 was used with the compound produced in theprevious step (500 mg, 2.46 mmol, 1.00 equiv) to afford 250 mg (99%) ofthe product as a colorless oil.

(2S)-2-[[(tert-Butoxy)carbonyl]amino]-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid

The method used to prepare 204 was used with(2S)-2-amino-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]pentanoicacid (1.29 g, 4.21 mmol, 1.00 equiv), 1,4-dioxane (50 mL), affording 1.3g (76%) of the product as a white solid.

tert-ButylN-[(2S)-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-[4-(d₃)-methylpiperazin-1-yl]-1-oxopentan-2-yl]carbamate

The method used to prepare 206 was used with the compound produced inthe previous step (1.075 g, 2.64 mmol, 1.00 equiv) and1-(d₃)-methylpiperazine (300 mg, 2.91 mmol, 1.10 equiv) to afford 650 mg(50%) of the product as a yellow oil.

(2S)-2-amino-5-[[(2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-[4-(d₃)-methylpiperazin-1-yl]pentan-1-one;trifluoroacetic acid salt

The method used to prepare 1 was used with the compound produced in theprevious step (650 mg, 1.32 mmol, 1.00 equiv), to afford 710 mg (96%) ofas a yellow oil.

N-[(2S)-5-[[(2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-[4-(d₃)-methylpiperazin-1-yl]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide

The method used to prepare 211 was used with the compound produced inthe previous step and 4-(1H-1,2,3-triazol-1-yl)benzoic acid (231 mg,1.22 mmol, 1.23 equiv) to afford 700 mg (96%) of the product as lightyellow oil.

N-[(2S)-5-[[(2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-[4-(d₃)-methylpiperazin-1-yl]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(137)

The method used to prepare 210 was used with the compound produced inthe previous step (850 mg, 1.51 mmol) to afford 112.2 mg (14%) of 137 asa light yellow solid.

Example 138

Synthesis of 138

1-(tert-Butoxycarbonyl)-(2,2,3,3,5,5,6,6-d₈)piperazine

Into a 250-mL round-bottom flask was added a solution of(2,2,3,3,5,5,6,6-d₈)piperazine dihydrochloride (1 g, 5.98 mmol, 1.00equiv) in methanol (10 mL), followed by the addition of a solution ofNaOH (480 mg, 12.00 mmol, 2.00 equiv) in methanol (10 mL). The mixturewas stirred for 30 min, then CF₃COOH (682 mg, 5.98 mmol, 1.00 equiv) wasadded. The reaction mixture was stirred for an additional 15 min, thenwater (20 mL) was added. The solution was stirred for 30 min, then asolution of Boc₂O (1.3 g, 5.96 mmol, 1.00 equiv) and I₂ (152 mg, 0.60mmol, 0.10 equiv) in methanol (40 mL) was added. The resulting solutionwas stirred for 3 h at 25° C., then concentrated under vacuum. The pHvalue of the solution was adjusted to 11 with NaOH (20%). The solidswere removed by filtration, and the resulting solution was extractedwith 3×30 mL of EtOAc. The organic layers were combined, washed with1×50 mL of brine, then dried over Na₂SO₄. The resulting solution wasconcentrated under vacuum to afford 1 g (86%) of the product asoff-white solid.

1-(tert-Butoxycarbonyl)-4-methyl-(2,2,3,3,5,5,6,6-d₈)piperazine

In a 100-mL round-bottom flask was combined the compound from theprevious step (1 g, 5.15 mmol, 1.00 equiv), THF (30 mL), and K₂CO₃ (2.14g, 15.48 mmol, 3.01 equiv). The mixture was stirred for 1 h, then asolution of CH₃I (730 mg, 5.14 mmol, 1.00 equiv) in THF (10 mL) wasadded dropwise with stirring at −12° C. The resulting solution wasstirred for 16 h at 25° C. The solids that formed were removed byfiltration. The resulting solution was concentrated under vacuum,diluted with 50 mL of H₂O, and extracted with 3×30 mL of EtOAc. Theorganic layers were combined, washed with 1×50 mL of brine, dried overNa₂SO₄, and concentrated under reduced pressure to afford 500 mg (47%)of the product as a light yellow oil.

1-Methyl-(2,2,3,3,5,5,6,6-d₈)piperazine

The procedure used to prepare 1 was used with the compound produced inthe previous step (500 mg, 2.40 mmol, 1.00 equiv) to afford 200 mg (37%)of PH-IMA-2013-003-336-3 as a light yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methyl-(2,2,3,3,5,5,6,6-d₈)-piperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)-benzamide

The method used to prepare 206 was used with the compound produced inthe previous step and(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]pentanoicacid (300 mg, 0.63 mmol, 1.00 equiv) to afford 150 mg (42%) of theproduct as a colorless oil.

N-[(2S)-5-[[(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amino]-1-(4-methyl-(2,2,3,3,5,5,6,6-d₈)-piperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)-benzamide(138)

The method used to prepare 210 was used with the compound produced inthe previous step (150 mg, 0.26 mmol) to afford 65.7 mg (47%) of 138 asa white solid.

Example 142

Synthesis of 142

(2S)-2-[(4-(1H-1,2,3-Triazol-1-yl)phenyl)formamido]-hexanedioic acid

In a 500-mL round-bottom flask were combined a solution of(2S)-2-aminohexanedioic acid (3 g, 18.62 mmol, 1.00 equiv) in water (50mL) and dioxane (50 mL), a solution of Na₂CO₃ (5.9 g, 55.67 mmol, 2.99equiv) in H₂O (50 mL). A solution of 4-(1H-1,2,3-triazol-1-yl)benzoylchloride (4.26 g, 20.52 mmol, 1.10 equiv) in dioxane (50 mL) was thenadded dropwise. The resulting solution was stirred for 1 h at 25° C. ThepH value of the solution was adjusted to 2 with HCl (2 M). The resultingsolution was extracted with 3×150 mL of EtOAc, and the organic layerswere combined, washed with 1×300 mL of brine, dried over Na₂SO₄,concentrated under vacuum, and applied onto a silica gel column withCH₂Cl₂/methanol to afford 4.5 g the product as a off-white solid.

(2S)-6-Methoxy-6-oxo-2-[(4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]-hexanoicacid

In a 100-mL round-bottom flask was added a solution of(2S)-2-[(4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]-hexanedioic acid(2500 mg, 7.52 mmol, 1.00 equiv) in methanol (40 mL), followed by thedropwise addition of AcCl (700 mg) with stirring at 0° C. The resultingsolution was stirred for 100 min at 0° C. The pH value of the solutionwas adjusted to 9 with sat. NaHCO₃. The resulting solution was extractedwith 2×20 mL of EtOAc and the aqueous layers were combined. HCl (2 M)was employed to adjust the pH to 2. The resulting solution was extractedwith 3×50 mL of EtOAc, and the organic layers were combined, washed with1×50 mL of brine, dried over Na₂SO₄, and concentrated under vacuum,affording 1.5 g (58%) of the product as a colorless oil.

Methyl(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]hexanoate

The method used to prepare 206 was used with the compound from theprevious step (1.5 g, 4.33 mmol, 1.00 equiv) and 1-methylpiperazine (500mg, 4.99 mmol, 1.73 equiv) to afford 1 g (54%) of the product as acolorless oil

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]hexanoicacid

The method used to prepare 205 was used with the compound from theprevious step (1 g, 2.33 mmol) to afford 950 mg (98%) of the product asa colorless oil

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]hexanol(212)

In a 100-mL round-bottom flask were combined the product from theprevious step (980 mg, 2.36 mmol, 1.00 equiv), N-methylmorpholine (500mg, 4.94 mmol, 2.09 equiv), and THF (50 mL), followed by the addition ofi-BuOCOCl (500 mg, 3.65 mmol, 1.54 equiv) dropwise with stirring at −20°C. The mixture was stirred for 2 h at −20° C. To this was added asolution of NaBH₄ (1 g, 26.43 mmol, 11.18 equiv) in methanol (20 mL)dropwise with stirring at −20° C. The resulting solution was stirred for2 h at 25° C., concentrated under vacuum, and applied onto a silica gelcolumn with CH₂Cl₂/methanol (5:1), to afford 300 mg (32%) of the productas a colorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(1H-1,2,3-triazol-1-yl)phenyl)formamido]hexanal

The method used to prepare 208 was used with the compound from theprevious step (300 mg, 0.75 mmol, 1.00 equiv) to afford 200 mg (67%) ofthe product as off-white solid.

N-[(2S)-6-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxohexan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(142)

The method used to prepare 4 was used with the compound from theprevious step (200 mg, 0.50 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (76 mg, 0.50 mmol, 1.00equiv) to afford 21.4 mg (7%) of 142 as a colorless oil.

Example 143

Synthesis of 143

(2S)-2-[(4-Cyanophenyl)formamido]-hexanedioic acid

The method used to prepare 203 was used with (2S)-2-aminohexanedioicacid (5 g, 31.03 mmol, 1.00 equiv) and 4-cyanobenzoyl chloride (5.2 g,31.41 mmol, 1.01 equiv) to afford 7 g (78%) of the product as acolorless oil.

(2S)-2-[(4-Cyanophenyl)formamido]-6-hydroxyhexanoic acid

In a 250-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen were combined the product from the previousstep (2 g, 6.89 mmol) and THF (100 mL), followed by the dropwiseaddition of BH₃ (17 mL, 1 M in THF) with stirring at 0° C. The resultingsolution was stirred for 2 h at 0° C. The reaction was then quenched bythe addition of 5 mL of methanol. The resulting mixture was concentratedunder reduced pressure, diluted with 50 mL of sat Na₂CO₃, and washedwith 3×30 mL of EtOAc. The aqueous layers were combined and adjusted topH 2 with HCl (2 M). The resulting solution was extracted with 3×50 mLof EtOAc, and the organic layers were combined, washed with 1×50 mL ofbrine, dried over Na₂SO₄, and concentrated under reduced pressure,affording 1 g (53%) of the product as a colorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[(4-cyanophenyl)formamido]hexanol

The method used to prepare 206 was used with(2S)-2-[4-cyanophenyl)formamido]-6-hydroxyhexanoic acid (1 g, 3.62 mmol,1.00 equiv) and 1-methylpiperazine (540 mg, 5.39 mmol, 1.49 equiv) toafford 1 g (77%) of the product as a colorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[(4-cyanophenyl)formamido]hexanal

The method used to prepare 208 was used with(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[(4-cyanophenyl)formamido]hexanol(500 mg, 1.39 mmol) to afford 450 mg (91%) of the product as off-whitesolid

N-[(2S)-6-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxohexan-2-yl]-4-cyanobenzamide(142)

The method used to prepare 4 was used with(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[(4-cyanophenyl)formamido]hexanal(450 mg, 1.26 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (250 mg, 1.65 mmol, 1.31equiv) to afford 174.1 mg (28%) of the product as a white solid.

Example 144

Synthesis of 144

(2S)-2-Amino-6-methoxy-6-oxo-hexanoic acid

In a 250-mL round-bottom flask were combined a solution of thionylchloride (6.2 mL) in methanol (40 mL) and (2S)-2-aminohexanedioic acid(10 g, 62.05 mmol, 1.00 equiv). The resulting solution was stirred for10 min at 0° C. in a water/ice bath. The reaction mixture was thenpoured into Et₂O (200 mL) and stirred for 10 min. The solid that formedwas collected by filtration, affording 10 g (92%) of the product as awhite solid

(2S)-6-Methoxy-6-oxo-2-[(4-(pyrimidin-2-yl)phenyl)formamido]-hexanoicacid

The method used to prepare 203 was used with(2S)-2-amino-6-methoxy-6-oxo-hexanoic acid (3 g, 14.17 mmol, 1.00 equiv)and 4-(pyrimidin-2-yl)benzoyl chloride (3 g, 13.72 mmol, 0.97 equiv) toafford 3 g (59%) of the product as an off-white solid.

Methyl(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanoate

The method used to prepare 206 was used with(2S)-6-methoxy-6-oxo-2-[(4-(pyrimidin-2-yl)phenyl)formamido]-hexanoicacid (3 g, 8.39 mmol, 1.00 equiv) and 1-methylpiperazine (1.3 g, 12.98mmol, 1.55 equiv) to afford 1 g (27%) of the product as a colorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanoicacid

The method used to prepare 205 was used with methyl(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanoate(1 g, 2.28 mmol, 1.00 equiv) to afford 600 mg (62%) of the product as acolorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanol

The method used to prepare 212 was used with(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanoicacid (600 mg, 1.41 mmol, 1.00 equiv) to afford 300 mg (52%) of theproduct as a colorless oil.

(5S)-6-(4-Methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanal

The method used to prepare 208 was used with(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanol(300 mg, 0.73 mmol) to afford 150 mg (50%) of the product as off-whitesolid.

N-[(2S)-6-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxohexan-2-yl]-4-(pyrimidin-2-yl)benzamide(144)

The method used to prepare 4 was used with(5S)-6-(4-methylpiperazin-1-yl)-6-oxo-5-[4-(pyrimidin-2-yl)phenyl)formamido]hexanal(150 mg, 0.37 mmol, 1.00 equiv) and(1S,2R)-2-(4-fluorophenyl)cyclopropan-1-amine (80 mg, 0.53 mmol, 1.44equiv) to afford 9.2 mg (5%) of 144 as a light yellow semi-solid.

Example 145

Synthesis of 145

(2R)-2-[(4-Fluorophenyl)formamido]-3-[(2-hydroxyethyl)sulfanyl]propanoicacid

In a 250-mL round-bottom flask was placed a solution of(2R)-2-[(4-fluorophenyl)formamido]-3-mercaptopropanoic acid (5 g, 20.55mmol) in N,N-dimethylformamide (50 mL) and K₂CO₃ (5.7 g, 40.94 mmol),followed by the addition of a solution of 2-bromoethanol (2.8 g, 22.41mmol) in N,N-dimethylformamide (20 mL) dropwise with stirring at 0° C.The resulting solution was stirred for 5 h at room temperature, thendiluted with 200 mL of H₂O. The pH of the solution was adjusted to 3with HCl (2 M). The resulting solution was extracted with 2×200 mL ofEtOAc, and the organic layers were combined, dried over Na₂SO₄ andconcentrated under vacuum. The residue was applied onto a silica gelcolumn with CH₂Cl₂/methanol (20:1), affording 4 g (68%) of the productas a yellow oil.

4-Fluoro-N-[(2R)-3-[(2-hydroxyethyl)sulfanyl]-1-(morpholin-4-yl)-1-oxopropan-2-yl]benzamide

In a 250-mL round-bottom flask were combined a solution of(2R)-2-[(4-fluorophenyl)formamido]-3-[(2-hydroxyethyl)sulfanyl]propanoicacid (4 g, 13.92 mmol, 1.00 equiv) in THF (50 mL), DEPBT (6.25 g, 20.90mmol, 1.50 equiv) and imidazole (1.42 g, 20.88 mmol, 1.50 equiv). Themixture was stirred for 30 min at 0° C., then morpholine (1.2 g, 13.77mmol, 0.99 equiv) was added. The resulting solution was stirred for 12 hat room temperature, diluted with 200 mL of EtOAc, and washed with 1×100mL of brine. The organic layers were dried over Na₂SO₄ and concentratedunder vacuum. The residue was applied onto a silica gel column withEtOAc/petroleum ether (1:10), affording 3 g (60%) of the product as ayellow oil.

N-[(2R)-3-([2-[tert-Butyldimethylsilyl)oxy]ethyl]sulfanyl)-1-(morpholin-4-yl)-1-oxopropan-2-yl]-4-fluorobenzamide

In a 250-mL round-bottom flask were combined a solution of4-fluoro-N-[(2R)-3-[(2-hydroxyethyl)sulfanyl]-1-(morpholin-4-yl)-1-oxopropan-2-yl]benzamide(3 g, 8.42 mmol, 1.00 equiv) in CH₂Cl₂ (30 mL) and imidazole (1.14 g,16.76 mmol, 1.99 equiv). followed by the addition of TBSCl (1.9 g, 12.58mmol, 1.49 equiv), dropwise at 0° C. The resulting solution was stirredfor 6 h at room temperature. The reaction was then quenched by theaddition of 50 mL of water. The resulting solution was extracted with2×100 mL of CH₂Cl₂ and the organic layers were combined and dried overanhydrous Na₂SO₄ and concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate/petroleum ether (1:30). Thisresulted in 2 g (50%) of the product as a white solid.

N-[(2R)-3-([2-[(tert-Butyldimethylsilyl)oxy]ethyl]sulfonyl)-1-(morpholin-4-yl)-1-oxopropan-2-yl]-4-fluorobenzamide

In a 250-mL round-bottom flask were combined a solution ofN-[(2R)-3-([2-[(tert-butyldimethylsilyl)oxy]ethyl]sulfanyl)-1-(morpholin-4-yl)-1-oxopropan-2-yl]-4-fluorobenzamide(2 g, 4.25 mmol, 1.00 equiv) in CH₂Cl₂ (20 mL) and m-CPBA (1.84 g, 10.66mmol, 2.51 equiv). The resulting solution was stirred for 6 h at roomtemperature, then diluted with 50 mL of CH₂Cl₂. The resulting mixturewas washed with 1×50 mL of saturated Na₂CO₃, then with 1×50 mL of brine.The organic layers were dried over anhydrous Na₂SO₄ and concentratedunder vacuum. The residue was applied onto a silica gel column withEtOAc/petroleum ether (1:30), affording 1.3 g (61%) of the product as awhite solid.

4-Fluoro-N-[(2R)-3-[(2-hydroxyethane)sulfonyl]-1-(morpholin-4-yl)-1-oxopropan-2-yl]benzamide

In a 50-mL round-bottom flask were combined a solution ofN-[(2R)-3-([2-[(tert-butyldimethylsilyl)oxy]ethyl]sulfonyl)-1-(morpholin-4-yl)-1-oxopropan-2-yl]-4-fluorobenzamide(500 mg, 0.99 mmol, 1.00 equiv) in THF (10 mL) and Bu₄NF (2 M) (1.5 mL).The resulting solution was stirred for 10 h at room temperature, thendiluted with 50 mL of EtOAc. The resulting mixture was washed with 2×10mL of brine, dried over anhydrous Na₂SO₄, and concentrated under vacuum.The residue was applied onto a silica gel column with EtOAc/petroleumether (1:20), affording 300 mg (78%) of the product as a yellow oil.

2-[[(2R)-2-[(4-Fluorophenyl)formamido]-3-(morpholin-4-yl)-3-oxopropyl]sulfonyl]ethylmethanesulfonate

In a 50-mL round-bottom flask were combined a solution of4-fluoro-N-[(2R)-3-[(2-hydroxyethyl)sulfonyl]-1-(morpholin-4-yl)-1-oxopropan-2-yl]benzamide(300 mg, 0.77 mmol, 1.00 equiv) in THF (5 mL) and Et₃N (156 mg, 1.54mmol, 2.00 equiv). This was followed by the addition of methanesulfonylchloride (134 mg, 1.17 mmol, 1.51 equiv) dropwise with stirring at 0° C.The resulting solution was stirred for 2 h at room temperature. Thereaction was then quenched by the addition of 10 mL of water. Theresulting solution was extracted with 3×10 mL of ethyl acetate. Thecombined organic layers were dried over Na₂SO₄ and concentrated undervacuum. The residue was applied onto a silica gel column withEtOAc/petroleum ether (1:50), affording 200 mg (56%) of the product as awhite solid.

Ethyl (1R)-2-(4-fluorophenyl)-1-methylcyclopropane-1-carboxylate

In a 100-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen were combined a solution of ethyl2-(diethoxyphosphoryl)propanoate (3.45 g, 14.48 mmol, 2.00 equiv) inethylene glycol dimethyl ether (20 mL), followed by the addition ofn-BuLi (2.5 M) (5.8 mL) dropwise with stirring at 0° C. The resultingsolution was stirred for 30 min at room temperature. To this was added2-(4-fluorophenyl)oxirane (1 g, 7.24 mmol, 1.00 equiv). The resultingsolution was allowed to react, with stirring, for an additional 12 hwhile the temperature was maintained at 80° C. in an oil bath. Thereaction mixture was cooled to room temperature and then quenched by theaddition of 20 mL of water. The resulting solution was extracted with3×50 mL of EtOAc, and the organic layers were combined, dried overanhydrous Na₂SO₄, concentrated under vacuum. The residue was appliedonto a silica gel column with EtOAc/petroleum ether (1:100), affording 1g (62%) of the product as a yellow oil.

(1R)-2-(4-Fluorophenyl)-1-methylcyclopropane-1-carboxylic acid

In a 50-mL round-bottom flask were combined a solution of ethyl(1R)-2-(4-fluorophenyl)-1-methylcyclopropane-1-carboxylate (1 g, 4.50mmol, 1.00 equiv) in methanol/H₂O (10/2 mL) and KOH (1.26 g, 22.46 mmol,4.99 equiv). The resulting solution was stirred for 10 h at roomtemperature, and then diluted with 20 mL of H₂O. The pH value of thesolution was adjusted to 2 with HCl (2 M). The resulting solution wasextracted with 3×20 mL of EtOAc, and the organic layers were combined,dried over anhydrous Na₂SO₄, and concentrated under vacuum, affording800 mg (92%) of the product as a yellow oil.

tert-Butyl N-[(1R)-2-(4-fluorophenyl)-1-methylcyclopropyl]carbamate

In a 50-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen were combined a solution of(1R)-2-(4-fluorophenyl)-1-methylcyclopropane-1-carboxylic acid (400 mg,2.06 mmol, 1.00 equiv) in toluene (10 mL), diphenylphosphoryl azide (680mg, 2.47 mmol, 1.20 equiv), and Et₃N (312 mg, 3.08 mmol, 1.50 equiv).The resulting solution was stirred for 30 min at 90° C. in an oil bath.tert-Butanol (2 mL) was then added. The resulting solution was allowedto react, with stirring, for an additional 12 h while the temperaturewas maintained at 90° C. in an oil bath. The reaction mixture was cooledto room temperature, then diluted with 50 mL of EtOAc. The resultingmixture was washed with 30 mL of H₂O, dried over anhydrous Na₂SO₄, thenconcentrated under vacuum. The residue was applied onto a silica gelcolumn with EtOAc/petroleum ether (1:100), affording 350 mg (64%) of theproduct as a yellow oil.

(1R,2S)-2-(4-Fluorophenyl)-1-methylcyclopropan-1-amine

Into a 50-mL round-bottom flask was added a solution of tert-butylN-[(1R,2S)-2-(4-fluorophenyl)-1-methylcyclopropyl]carbamate (350 mg,1.32 mmol, 1.00 equiv) in methanol(HCl) (10 mL). The resulting solutionwas stirred for 2 h at room temperature, then diluted with 10 mL of H₂O.The pH value of the solution was adjusted to 9 with saturated Na₂CO₃.The resulting solution was extracted with 3×10 mL of EtOAc, and theorganic layers were combined and dried over anhydrous Na₂SO₄ andconcentrated under vacuum, affording 200 mg (92%) of the product as ayellow oil.

4-Fluoro-N-[(2R)-3-[(2-[[(1R,2S)-2-(4-fluorophenyl)-1-methylcyclopropyl]amino]ethane)sulfonyl]-1-(morpholin-4-yl)-1-oxopropan-2-yl]benzamide(145)

In a 25-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined a solution of2-[[(2R)-2-[(4-fluorophenyl)formamido]-3-(morpholin-4-yl)-3-oxopropane]sulfonyl]ethylmethanesulfonate (200 mg, 0.43 mmol, 1.00 equiv) in MeCN (10 mL),i-Pr₂NEt, (110 mg, 0.85 mmol, 1.99 equiv), KI (71 mg, 0.43 mmol, 1.00equiv), and (1R,2S)-2-(4-fluorophenyl)-1-methylcyclopropan-1-amine (71mg, 0.43 mmol, 1.00 equiv). The resulting solution was stirred for 12 hat 50° C. in an oil bath, and then diluted with 30 mL of EtOAc. Theresulting mixture was washed with 1×20 mL of H₂O, dried over Na₂SO₄,then concentrated under vacuum. The crude product was purified by HPLC,affording 64.3 mg (28%) of the product as a white solid.

Example 146

Synthesis of 146

4-(3,5-dimethylpyrazol-1-yl)benzoic acid

In a 250-mL round-bottom flask were combined 4-hydrazinylbenzoic acid(8.8 g, 57.84 mmol, 1.00 equiv), AcOH (100 mL), and pentane-2,4-dione(5.8 g, 57.93 mmol, 1.00 equiv). The resulting solution was stirred for16 h at 118° C. in an oil bath, and concentrated under vacuum. Theresulting solid was washed with 3×100 mL of EtOAc and dried undervacuum, affording 9.2 g (74%) of the product as a light-brown solid.

(2S)-2-[(4-(3,5-dimethylpyrazol-1-yl)phenyl)formamido]-4-[(tert-butyldiphenylsilyl)oxy]butanoicacid

The method used to prepare 211 was used with the compound from theprevious step (5 g, 23.12 mmol, 1.50 equiv) and(2S)-2-amino-4-[(tert-butyldiphenylsilyl)oxy]butanoic acid (5.52 g,15.44 mmol, 1.00 equiv) to afford 1 g (12%) of the product as a yellowsolid.

(2S)—N-[1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxo-4-((tert-butyldiphenylsilyl)oxy)-butan-2-yl]-4-(4-(3,5-dimethylpyrazol-1-yl)benzamide

The method used to prepare 206 was used with the compound produced inthe previous step (5 g, 9.00 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide (1.46 g, 10.8 mmol, 1.20 equiv) to afford 1 g(17%) of the product as a yellow oil.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[[4-(3,5-dimethylpyrazol-1-yl)phenyl]formamido]-butanol

The method used to prepare 207 was used with the compound from theprevious step (1 g, 1.49 mmol, 1.00 equiv) to afford 300 mg (46%) of theproduct as a yellow oil.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[[4-(3,5-dimethylpyrazol-1-yl)phenyl]formamido]-butanal

The method used to prepare 208 was used with the compound from theprevious step (50 mg, 0.11 mmol, 1.00 equiv) to afford 45 mg (91%) ofthe product as a light yellow solid.

N-[(2S)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl]-4-(3,5-dimethylpyrazol-1-yl)benzamide(146)

The method used to prepare 4 was used with the compound from theprevious step (45 mg, 0.10 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (19 mg, 0.125 mmol, 1.20equiv) to afford 11.7 mg (20%) of 146 as a white solid.

Example 148

Synthesis of 148

(2S)-4-[(tert-Butyldiphenylsilyl)oxy]-2-[[(9H-fluoren-9-ylmethoxy)carbonyl]amino]butanoicacid

In a 1000-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was combined a solution of(2S)-2-amino-4-[(tert-butyldiphenylsilyl)oxy]butanoic acid (10 g, 27.97mmol, 1.00 equiv) in MeCN (300 mL), 5% aqueous NaHCO₃ (300 mL), andFmoc-OSu (10.4 g, 30.86 mmol, 1.10 equiv). The resulting solution wasstirred for 12 h at room temperature. The pH value of the solution wasadjusted to 3 with HCl (2 M). The resulting solution was extracted with3×200 mL of EtOAc. The organic layers were combined, dried over Na₂SO₄,concentrated under reduced pressure, and applied onto a silica gelcolumn with EtOAc/petroleum ether (1:1), to afford 11 g (68%) of theproduct as a yellow solid.

9H-Fluoren-9-ylmethylN-[(2S)-4-[(tert-butyldiphenylsilyl)oxy]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl]carbamate

The procedure of 206 was used with the compound from the previous stepand thiomorpholine-1,1-dioxide (3.9 g, 22.59 mmol, 1.19 equiv) to afford9 g (68%) of the product as a yellow solid.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[((9H-fluoren-9-ylmethyl)oxycarbonyl)amino]-1-butanol

The method used to prepare 207 was used with the compound from theprevious step (5 g, 7.17 mmol, 1.00 equiv) to afford 1.7 g of theproduct as an off-white solid.

(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[((9H-fluoren-9-ylmethyl)oxycarbonyl)amino]-1-butanal

The method used to prepare 208 was used with the compound from theprevious step (1.7 g, 3.71 mmol) to afford 1.2 g (71%) of the product asa yellow solid.

9H-fluoren-9-ylmethylN-[(2R)-1-(thiomorpholine-1,1-dioxide-4-yl)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxobutan-2-yl]carbamate

The method used to prepare 4 was used with the compound from theprevious step (1.2 g, 2.63 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (476 mg, 3.15 mmol, 1.20equiv) to afford 1.4 g (90%) of the product as a yellow solid.

tert-ButylN-[(3R)-4-(thiomorpholine-1,1-dioxide-4-yl)-3-[[(9H-fluoren-9-ylmethoxy)carbonyl]amino]-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate

The method used to prepare 204 was used with the product from theprevious reaction (1.4 g, 2.37 mmol, 1.00 equiv) to afford 1.5 g (92%)of the product as a yellow solid.

tert-ButylN-[(3S)-3-amino-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate

In a 100-mL round-bottom flask was combined the compound from theprevious step (1.5 g, 2.17 mmol, 1.00 equiv), piperidine (5 mL), and DMF(20 mL). The resulting solution was stirred for 12 h at roomtemperature, then diluted with 100 mL of H₂O. The solids that formedwere removed by filtration. The resulting solution was extracted with3×20 mL of EtOAc, and the organic layers were combined, dried overNa₂SO₄, and concentrated under reduced pressure, affording 800 mg (79%)of the product as a yellow oil.

tert-ButylN-[(3S)-3-[[4-(2,5-dimethyl-1H-pyrrol-1-yl)phenyl]formamido]-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate

The method used to prepare 211 was used with the product from theprevious step (397 mg, 0.85 mmol, 1.00 equiv) and4-(2,5-dimethyl-1H-pyrrol-1-yl)benzoic acid (200 mg, 0.93 mmol, 1.10equiv) to afford 200 mg (35%) of the product as a yellow oil.

4-(2,5-Dimethyl-1H-pyrrol-1-yl)-N-[(2S)-1-(thiomorpholine-1,1-dioxide-4-yl)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxobutan-2-yl]benzamide,trifluoroacetic acid salt

The method used to prepare 1 was used with the product from the previousstep (120 mg, 0.18 mmol, 1.00 equiv) to afford 12 mg (10%) of theproduct as a light yellow solid.

Example 150

(1R,2S)-2-(4-fluorophenyl)cyclopropanecarboxylic acid

Into a 250-mL round-bottom flask, was placed a solution of(1R)—N-(2-hydroxy-1-phenylethyl)-2-(4-fluorophenyl)-1-methylcyclopropane-1-carboxamide(5 g, 16.70 mmol, 1.00 equiv) in dioxane (100 ml) and H₂SO₄ (30 mL). Theresulting solution was stirred for 16 h at 100° C. in an oil bath. Theresulting solution was then diluted with 300 mL of CH₂Cl₂ The combinedorganic layers were washed with 2×200 mL of H₂O and 1×500 mL of brine,dried over Na₂SO₄, and concentrated under reduced pressure to afford 2 g(66%) of the product as a colorless oil.

N-(tert-butoxycarbonyl)-(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amine

In a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was combined a solution of the compound from theabove step (2 g, 11.10 mmol, 1.00 equiv) in toluene (100 mL). Then DPPA(4.6 g, 16.72 mmol, 1.51 equiv) and Et₃N (1.7 g, 16.80 mmol, 1.51 equiv)were added at room temperature. The reaction mixture was stirred at 110°C. in an oil bath. After 30 min, the reaction mixture was cooled to 90°C. Then t-BuOH (20 mL) was added to the solution. The resulting solutionwas stirred for 5 h at 90° C., then extracted with 3×100 mL of EtOAc.The combined organic layers were washed with brine, dried over Na₂SO₄,concentrated under reduced pressure, and applied onto a silica gelcolumn with EtOAc/petroleum ether (1:10) to afford 2 g (72%) of theproduct as a white solid.

(1R,2S)-2-(4-Fluorophenyl)cyclopropyl]amine

In a 100-mL round-bottom flask were combined a solution of the compoundfrom the previous step (2 g, 7.93 mmol) and HCl/MeOH (50 mL). Theresulting solution was stirred for 1 h at room temperature. The reactionmixture was then concentrated under reduced pressure, affording 1.4 g(92%) of the product as a colorless oil.

N-[(2S)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxobutan-2-yl]-4-phenylbenzamide

The method used to prepare 4 was used with the product from the abovereaction (137 mg, 0.72 mmol, 1.50 equiv) and(3S)-4-(thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[4-(phenyl)phenyl)formamido]-1-butanal(200 mg, 0.48 mmol, 1.00 equiv) to afford 106.9 mg (34.4%) of theproduct as a white solid.

Example 152

Synthesis of 152

tert-ButylN-[(3S)-4-(thiomorpholin-1,1-dioxide-4-yl)-3-[[4-(4-methanesulfonylphenyl)phenyl]formamido]-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen were combined tert-butylN-[(3S)-3-amino-4-(thiomorpholin-1,1-dioxide-4-yl)-4-oxobutyl]-N-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]-carbamate(100 mg, 0.21 mmol, 1.00 equiv), Et₃N (43 mg, 0.42 mmol, 2.00 equiv),CH₂Cl₂ (10 mL), followed by the dropwise addition of a solution of4-(4-methanesulfonylphenyl)benzoyl chloride (70 mg, 0.24 mmol, 1.12equiv) in CH₂Cl₂ (5 mL) dropwise with stirring. The resulting solutionwas stirred for 2 h at room temperature, then quenched by the additionof 10 mL of water. The resulting solution was extracted with 3×10 mL ofCH₂Cl₂. The combined organic layers were dried Na₂SO₄, concentratedunder reduced pressure, and applied onto a silica gel column withEtOAc/petroleum ether (1:5) to afford 100 mg (65%) of the product as ayellow oil.

N-[(2S)-1-(thiomorpholin-1,1-dioxide-4-yl)-4-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxobutan-2-yl]-4-(4-methanesulfonylphenyl)benzamidetrifluoroacetic acid salt

In a 50-mL round-bottom flask were combined the compound from theprevious step (100 mg, 0.14 mmol, 1.00 equiv), CF₃COOH (1 mL) and CH₂Cl₂(10 mL). The resulting solution was stirred for 2 h at room temperature,then concentrated under reduced pressure. The crude product was purifiedby Prep-HPLC, affording 51.5 mg (52%) of the produce as a white solid.

Example 158

Synthesis of 158

4-(1H-1,2,3-triazolyl-1-yl)benzoyl chloride (1)

In a 100-mL round-bottom flask were combined4-(1H-1,2,3-triazol-1-yl)benzoic acid (1 g, 5.29 mmol, 1.00 equiv) andthionyl chloride (20 mL). The resulting solution was stirred for 16 h at80° C. in an oil bath. The resulting mixture was then concentrated underreduced pressure, affording 1 g (91%) of intermediate (1) as a yellowsolid.

(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](propen-3-yl)amino]-2-[[4-(1H-1,2,3-triazol-1-yl)phenyl]formamido]pentanoicacid (2)

In a 100-mL round-bottom flask were combined(2S)-2-amino-5-[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)aminopentanoicacid (500 mg, 1.63 mmol, 1.00 equiv), Et₃N (494 mg, 4.88 mmol, 3.00equiv) and THF (20 mL). This was followed by the addition of a solutionof intermediate (1) from the previous step (1 g, 4.82 mmol, 2.95 equiv)in THF (20 mL) dropwise with stirring at 0° C. in 30 min. The resultingsolution was stirred for 1 h at 0° C. in an ice/salt bath, thenconcentrated under reduced pressure, and applied onto a silica gelcolumn with CH₂Cl₂/methanol (10:1). The collected fractions werecombined and concentrated under reduced pressure, affording 400 mg (51%)of intermediate (2) as a off-white solid.

N-[(2S)-1-(4-(methyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)-cyclopropyl](prop-2-en-1-yl)amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(3)

In a 100-mL round-bottom flask were combined intermediate (2) from theprevious step (400 mg, 0.84 mmol, 1.00 equiv), DEPBT (375 mg, 1.25 mmol,1.50 equiv), and THF (20 mL), followed by the addition of imidazole (85mg, 1.25 mmol, 1.50 equiv). The mixture was stirred for 30 min at 0° C.,at which point 1-methylpiperazine (127 mg, 1.27 mmol, 1.50 equiv) wasadded dropwise with stirring at 0° C. in 3 min. The resulting solutionwas stirred for 16 h at 20° C., then concentrated under reducedpressure. The residue was applied onto a silica gel column withCH₂Cl₂/methanol (10:1). The collected fractions were combined andconcentrated under vacuum, affording 300 mg (64%) of intermediate (3) asa yellow solid.

N-[(2S)-1-(4-(methyl)piperazin-1-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)-cyclopropyl]amino]-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(3)

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placedN-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(1H-1,2,3-triazol-1-yl)benzamide(300 mg, 0.54 mmol, 1.00 equiv), 1,3-dimethyl-1,3-diazinane-2,4,6-trione(210 mg, 1.34 mmol, 2.50 equiv), Pd(PPh₃)₄ (155 mg, 0.13 mmol, 0.25equiv). The resulting solution was stirred for 2 h at 45° C. in an oilbath. The resulting mixture was concentrated under vacuum. The crudeproduct (10 mL) was purified by Flash-Prep-HPLC. This resulted in 65 mg(23%) of Example 158 as a yellow solid.

Alternatively, Example 158 and its bis-tosylate salt may be prepared bythe following method.

Example 162

Synthesis of 162

Methyl 4-(pyridin-2-yl)benzoate

In a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was combined a solution of methyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (5 g, 0.019 mol,1.00 equiv) in dioxane (50 mL), Na₂CO₃ (6.04 g, 0.057 mol, 3.00 equiv),2-bromopyridine (4.5 g, 0.028 mol, 1.50 equiv), and Pd(Ph₃P)₄ (1.1 g,0.001 mmol, 0.05 equiv). The resulting solution was stirred for 16 h at80° C. The resulting solution was quenched by water/ice and extractedwith 3×50 ml EtOAc. The combined organic layers were washed with brine,dried over Na₂SO₄, concentrated under reduced pressure, and purified byflash chromatography with EtOAc/petroleum ether (1:5) to afford 3.5 g(86%) of the product as a light yellow solid.

4-(Pyridin-2-yl)benzoic acid

The method used to prepare 209 was used with the compound from theprevious step (3.5 g, 0.016 mol, 1.00 equiv) in MeOH (50 mL) to afford 3g (92%) of the product as a off-white solid.

4-(Pyridin-2-yl)benzoyl chloride

The method used to prepare 201 was used with the compound from theprevious reaction (3 g, 0.015 mol, 1.00 equiv). The crude product wasused for the next step without further purification.

(S)-5-Methoxy-5-oxo-2-(4-(pyridin-2-yl)benzamido)pentanoic acid

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of(2S)-2-amino-5-methoxy-5-oxopentanoic acid (2 g, 12.41 mmol, 1.00 equiv)in dioxane (25 mL), a solution of Na₂CO₃ (3.94 g, 37.17 mmol, 3.00equiv) in water (50 mL). Then 4-(pyridin-2-yl)benzoyl chloride (crude)in N-methyl-2-pyrrolidone (100 ml) was added dropwise at 0° C. Theresulting solution was stirred for 1 h at 0° C. The pH value of thesolution was adjusted to 6 with HCl (1 M). The resulting mixture wasconcentrated under reduced pressure. The residue was purified by reversephase flash chromatography with CH₃CN/H₂O (1%-20%), affording 1.9 g(45%) of the product as a light yellow solid.

Methyl(4S)-5-oxo-5-[1-(thiomorpholine-1,1-dioxide-4-yl)]-4-[(4-(2-pyridyl)phenyl)formamido]pentanoate

The method used to prepare 206 was used with the product from theprevious step (1.9 g, 5.56 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide (1.13 g, 8.34 mmol, 1.50 equiv), to afford1.4 g (55%) of the product as a brown solid.

(4S)-5-Oxo-5-[1-(thiomorpholine-1,1-dioxide-4-yl)]-4-[(4-(2-pyridyl)phenyl)formamido]pentanoicacid

The procedure used to prepare 205 was used with the compound from theprevious step (1.4 g, 3.04 mmol, 1.00 equiv) to afford 1.2 g (89%) ofthe product as an off-white solid.

(4S)-5-Oxo-5-[1-(thiomorpholine-1,1-dioxide-4-yl)]-4-[(4-(2-pyridyl)phenyl)formamido]pentanol

The method used to prepare 212 was used with the product from theprevious step (800 mg, 1.8 mmol, 1.00 equiv) to afford 550 mg (71%) ofthe product as a brown solid.

(4S)-5-Oxo-5-[1-(thiomorpholine-1,1-dioxide-4-yl)]-4-[(4-(2-pyridyl)phenyl)formamido]pentanal

The method used to prepare 208 was used with the compound from theprevious step (550 mg, 1.27 mmol, 1.00 equiv) to afford 350 mg (64%) ofthe product as a off-white solid.

[(2S)-1-(Thiomorpholine-1,1-dioxide-4-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)-cyclopropyl]amino]-1-oxopentan-2-yl]4-(pyridin-2-yl)-benzamide

The method used to prepare 4 was used with the compound from theprevious step (350 mg, 0.82 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine hydrochloride salt (231mg, 1.23 mmol, 1.50 equiv) to afford 61.9 mg (13.4%) of the product as awhite solid.

Example 163

Synthesis of 163

4-(4H-1,2,4-Triazol-4-yl)benzoyl chloride

The method used to prepare 201 was used with4-(4H-1,2,4-triazol-4-yl)benzoic acid (2 g, 10.57 mmol, 1.00 equiv) toafford 2 g (91%) of the product as a white solid.

(2S)-5-Methoxy-5-oxo-2-[[4-(4H-1,2,4-triazol-4-yl)phenyl]formamido]pentanoicacid

In a 500-mL round-bottom flask were combined(2S)-2-amino-5-methoxy-5-oxopentanoic acid (1.55 g, 9.62 mmol, 1.00equiv), dioxane (100 mL), water (100 mL), and Na₂CO₃ (3.06 g, 28.87mmol, 3.00 equiv), followed by the dropwise addition of a solution of4-(4H-1,2,4-triazol-4-yl)benzoyl chloride (2 g, 9.63 mmol, 1.00 equiv)in NMP (50 mL) dropwise with stirring at 0° C. The resulting solutionwas stirred for 2 h at room temperature, then concentrated under vacuumand purified by Prep-HPLC to afford 2 g (63%) of the product as a yellowsolid.

Methyl(4S)-5-(thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(4H-1,2,4-triazol-4-yl)phenyl]formamido]pentanoate

The method used to prepare 206 was used with the product from theprevious step (2 g, 6.02 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide (1.22 g, 8.96 mmol, 1.49 equiv) to afford 1.4g (52%) of the product as a yellow oil.

(4S)-5-(Thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(4H-1,2,4-triazol-4-yl)phenyl]formamido]pentanoicacid

The method used to prepare 205 was used with the compound from theprevious step (1.4 g, 3.11 mmol, 1.00 equiv) to afford 1 g (74%) of theproduct as a white solid.

(4S)-5-(Thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(4H-1,2,4-triazol-4-yl)phenyl]formamido]pentanol

The method used to prepare 202 was used with the compound from theprevious step (900 mg, 2.07 mmol, 1.00 equiv) to afford 450 mg (52%) ofthe product as a yellow oil.

(4S)-5-(Thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(4H-1,2,4-triazol-4-yl)phenyl]formamido]pentanal

The method used to prepare 208 was used with the compound from theprevious step (450 mg, 1.07 mmol, 1.00 equiv) to afford 300 mg (67%) ofthe product as a yellow oil.

N-[(2S)-1-(Thiomorpholine-1,1-dioxide-4-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(4H-1,2,4-triazol-4-yl)benzamide

The method used to prepare 4 was used with the product from the previousstep (300 mg, 0.72 mmol, 1.00 equiv) to afford 17 mg (4%) of the productas an off-white solid.

Example 164

Synthesis of 164

4-(1H-1,2,3-Triazol-1-yl)-benzonitrile

In a 100-mL round-bottom flask was combined 4-fluorobenzonitrile (2 g,16.51 mmol, 1.00 equiv), Cs₂CO₃ (10.8 g, 2.00 equiv), 1H-1,2,3-triazole(1.4 g, 1.20 equiv), and N,N-dimethylformamide (50 mL). The resultingsolution was stirred for 4 h at 80° C., then diluted with 50 mL of H₂Oand extracted with 3×50 mL of EtOAc. The organic layers were combined,washed with 1×100 mL of brine, dried over Na₂SO₄, concentrated underreduced pressure, and applied silica gel column with EtOAc, affording1.2 g (43%) of the product as a yellow solid.

4-(1H-1,2,3-Triazol-1-yl)-benzoic acid

In a 100-mL round-bottom flask was dissolved the compound from theprevious reaction (1 g, 5.88 mmol, 1.00 equiv) in H₂SO₄ (20 mL, 9mol/L). The resulting solution was stirred for 16 h at 25° C. The solidsthat formed were collected by filtration, affording 1 g (90%) of theproduct as a gray solid.

4-(1H-1,2,3-Triazol-1-yl)-benzoyl chloride

In a 100-mL round-bottom flask were combined the compound from theprevious step (7 g, 37.04 mmol, 1.00 equiv) and thionyl chloride (30mL). The resulting solution was stirred for 16 h at 80° C. in an oilbath. The resulting mixture was concentrated under vacuum, affording 6 g(80%) of the product as a light yellow solid.

N-((2S)-5-((1R,2S)-2-(4-Fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-pentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamide

The remainder of the synthesis proceeded as for Scheme 18,

Example 166

Synthesis of 166

1-Phenyl-1H-pyrazole-4-carbonyl chloride hydrochloride

The method used to prepare 201 was used with1-phenyl-1H-pyrazole-4-carboxylic acid (5 g, 26.57 mmol, 1.00 equiv) toafford 5 g (77%) crude of the product as a yellow solid.

(2S)-5-Methoxy-5-oxo-2-[(1-phenyl-1H-pyrazol-4-yl)formamido]pentanoicacid

The method used to prepare 203 was used with(2S)-2-amino-5-methoxy-5-oxopentanoic acid (2.76 g, 17.13 mmol, 1.00equiv) and 1-phenyl-1H-pyrazole-4-carbonyl chloride hydrochloride (5 g,20.57 mmol, 1.20 equiv) to afford 2.7 g (48%) of the product as anoff-white solid.

Methyl(4S)-5-(thiomorpholin-1,1-dioxide-4-yl)-5-oxo-4-[(1-phenyl-1H-pyrazol-4-yl)formamido]pentanoate

The method used to prepare 206 was used with the compound from theprevious step (2.7 g, 8.15 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide (1.34 g, 9.84 mmol, 1.21 equiv) to afford 1.8g (49%) of the product as an off-white solid.

(4S)-5-(Thiomorpholin-1,1-dioxide-4-yl)-5-oxo-4-[(1-phenyl-1H-pyrazol-4-yl)formamido]pentanoicacid

The method used to prepare 205 was used with the compound from theprevious step (1.8 g, 4.01 mmol) to afford 1.22 g (70%) of the productas an off-white solid.

(3S)-4-(Thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[4-(1H-pyrazol-1-yl)phenyl)formamido]-1-butanol

The method to prepare 202 was used with the compound from the previousstep to afford 0.8 g (68%) of the product as an off-white solid.

(3S)-4-(Thiomorpholine-1,1-dioxide-4-yl)-4-oxo-3-[4-(1H-pyrazol-1-yl)phenyl)formamido]-1-butanal

The method used to prepare 208 was used with the compound from theprevious step (800 mg, 1.90 mmol, 1.00 equiv) to afford 480 mg (60%) ofthe product as a off-white solid.

N-[(2S)-1-(Thiomorpholine-1,1-dioxide-4-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-1-phenyl-1H-pyrazole-4-carboxamide

The method used to prepare 4 was used with the compound from theprevious reaction (480 mg, 1.15 mmol, 1.00 equiv) and(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (208 mg, 1.38 mmol, 1.20equiv) to afford 33.5 mg (5%) of the product as a white solid.

Example 168

Synthesis of 168

Methyl 4-(pyrimidin-2-yl)benzoate

In a 250-mL round-bottom flask were combined[4-(methoxycarbonyl)phenyl]boronic acid (5 g, 27.78 mmol, 1.00 equiv),dixoane (100 mL), 2-bromopyrimidine (5.3 g, 33.34 mmol, 1.20 equiv),Na₂CO₃ (5.89 g, 55.57 mmol, 2.00 equiv), water (10 mL), and Pd(Ph₃P)₂Cl₂(3.2 g, 2.77 mmol, 0.10 equiv). The resulting solution was stirred for16 h at 80° C. The solids that formed were removed by filtration. Theresulting solution was extracted with 3×50 mL of EtOAc. The organiclayers were combined and applied onto a silica gel column withCH₂Cl₂/methanol (10:1) to afford 4.4 g (74%) of the product as a lightyellow solid.

4-(Pyrimidin-2-yl)benzoic acid

In a 250-mL round-bottom flask were combined the compound from theprevious step (4.4 g, 20.54 mmol, 1.00 equiv), NaOH (2.4 g, 60.00 mmol,2.92 equiv), and methanol (50 mL). The resulting solution was stirredfor 2 h at room temperature. The pH value of the solution was adjustedto 7 with HCl (1 M). The solids that formed were collected byfiltration, affording 3.15 g (77%) of the product as a light yellowsolid.

4-(Pyrimidin-2-yl)benzoyl chloride

The method used to prepare 201 was used with the compound from theprevious step (3.15 g, 15.73 mmol, 1.00 equiv) affording 3.20 g (80%) ofcrude 4-(pyrimidin-2-yl)benzoyl chloride hydrochloride as a light yellowsolid.

(2S)-5-Methoxy-5-oxo-2-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanoicacid

The method used to prepare 203 was used with the compound from theprevious step (1.68 g, 10.42 mmol, 0.83 equiv) and4-(pyrimidin-2-yl)benzoyl chloride hydrochloride (3.20 g, 12.54 mmol,1.00 equiv) to afford 1.75 g (41%) of the product as a off-white solid.

Methyl(4S)-5-(thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanoate

The method used to prepare 206 was used with the compound from theprevious step (1.7 g, 4.95 mmol, 1.00 equiv) andthiomorpholine-1,1-dioxide (800 mg, 5.87 mmol, 1.19 equiv), to afford1.35 g (59%) of the product as a light yellow solid.

(4S)-5-(thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanoicacid

The method used to prepare 205 was used with the product from theprevious step (1.35 g, 2.93 mmol, 1.00 equiv) to afford 0.98 g (75%) ofthe product as a off-white solid.

(4S)-5-(thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanol

The method used to prepare 212 was used with the compound from theprevious step (980 mg, 2.19 mmol, 1.00 equiv), to afford 710 mg (75%) ofthe product as a solid.

(4S)-5-(thiomorpholine-1,1-dioxide-4-yl)-5-oxo-4-[[4-(pyrimidin-2-yl)phenyl]formamido]pentanal

The method used to prepare 208 was used with the compound from theprevious step (710 mg, 1.64 mmol, 1.00 equiv) to afford (72%) of theproduct as a white solid.

N-[(2S)-1(thiomorpholine-1,1-dioxide-4-yl)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide

The method that was used to prepare 4 was used with the compound fromthe previous step (510 mg, 1.18 mmol, 1.00 equiv) to afford 59.9 mg (9%)of the product as a off-white solid.

Example 180

Synthesis of 180

Ethyl 4-(1H-pyrrol-1-yl) benzoate

In a 250-mL round-bottom flask was combined a solution of ethyl4-aminobenzoate (4 g, 24.21 mmol, 1.00 equiv) in CH₂Cl₂ (100 mL), asolution of AcOH (300 mg, 5.00 mmol, 0.20 equiv) in water (30 mL), NaOAc(2 g, 24.39 mmol, 1.00 equiv), and 2,5-dimethoxyoxolane (4.8 g, 36.32mmol, 1.50 equiv). The resulting solution was stirred for 16 h at 90° C.in an oil bath. The resulting mixture was concentrated under vacuum,affording 3.5 g (67%) of the product as an off-white solid.

4-(1H-Pyrrol-1-yl) benzoic acid

In a 250-mL round-bottom flask was combined the compound from theprevious step (3.5 g, 16.26 mmol, 1.00 equiv), methanol (100 mL), andNaOH (1.3 g, 32.50 mmol, 2.00 equiv). The resulting solution was stirredfor 16 h at 50° C. in an oil bath. The pH value of the solution wasadjusted to 7 with HCl (1 M). The solids that formed were collected byfiltration, affording 3 g (99%) of the product as a white solid.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](propen-3-yl)amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxopentan-2-yl]-4-(1H-pyrrol-1-yl)benzamide

The procedure of 211 was used with the compound from the previous step(100 mg, 0.53 mmol, 1.50 equiv) and(2S)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(thiomorpholine-1,1-dioxide-4-yl)pentan-1-one(150 mg, 0.35 mmol, 1.00 equiv), affording 150 mg (72%) of the productas a yellow solid.

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(thiomorpholine-1,1-dioxide-4-yl)-1-oxopentan-2-yl]-4-(1H-pyrrol-1-yl)benzamide

The method to prepare 210 was used with the compound from the previousstep to afford 41.5 mg (30%) of the product as a light yellow solid.

Example 181

Into a 250-mL round-bottom flask, was placed(2S)-2-amino-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)pentan-1-one(1.04 g, 2.68 mmol, 1.00 equiv), dichloromethane (50 mL), HATU (1.71 g,4.50 mmol, 1.68 equiv), DIEA (1.16 g, 8.98 mmol, 3.35 equiv). This wasfollowed by the addition of a solution of 4-(pyrimidin-2-yl)benzoic acid(450 mg, 2.25 mmol, 0.84 equiv) in CH₂Cl₂ (5 mL) dropwise with stirringat 0° C. The resulting solution was stirred overnight at roomtemperature. The resulting mixture was concentrated under vacuum. Theresulting solution was extracted with 3×50 mL of EtOAc and the organiclayers combined. This resulted in 0.52 g (34%) ofN-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamideas a red solid.

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placedN-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl](prop-2-en-1-yl)amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamide(520 mg, 0.91 mmol, 1.00 equiv), THF (15 mL),1,3-dimethyl-1,3-diazinane-2,4,6-trione (426 mg, 2.73 mmol, 2.99 equiv),Pd(PPh₃)₄ (210 mg, 0.18 mmol, 0.20 equiv). The resulting solution wasstirred for 2 h at 50° C. in an oil bath. The solids were filtered out.The resulting solution was extracted with 3×50 mL of EtOAc and theorganic layers were combined and concentrated under vacuum. The mixturewas dried over Na₂SO₄. The crude product was purified by Prep-HPLC. Thisresulted in 180.9 mg (37%) ofN-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(pyrimidin-2-yl)benzamideas a light yellow solid.

Example 182

Synthesis of 182

(S)-methyl 1-tritylaziridine-2-carboxylate [2]

To a solution of (S)-methyl 3-hydroxy-2-(tritylamino)propanoate (1, 2.00g, 5.54 mmol) in chloroform (30 mL) was added with triethylamine (2.08mL, 14.95 mmol) and dimethylaminopyridine (67 mg, 0.55 mmol) followed bymethanesulfonyl chloride (888 mg, 7.75 mmol). The reaction mixture wasstirred under nitrogen atmosphere at 65° C. for 16 h. After this time,the reaction was cooled to room temperature and diluted with water (50mL). The layers were separated and the aqueous layer was extracted withethyl acetate (2×100 mL). The combined organic layers were washed withbrine (100 mL), dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The resulting residue was recrystallized fromethanol to afford 2 (1.20 g, 63%,) as an off-white solid.

(S)-methyl aziridine-2-carboxylate trifluoroacetic acid salt [3]

To a stirred solution of 2 (1.00 g, 2.91 mmol) in a mixture of methanol(3.0 mL) and chloroform (3.0 mL) at 0° C. was added trifluoroacetic acid(2.1 mL). The mixture was stirred under nitrogen atmosphere at roomtemperature for 2 h. After this time, the reaction mixture wasconcentrated under reduced pressure to afford crude 3 (600 mg) as acolorless liquid, which was used without further purification.

4-(1H-pyrazol-1-yl)benzoyl chloride[4]

To a solution of 4-(1H-pyrazol-1-yl)benzoic acid (400 mg, 2.12 mmol) inmethylene chloride (8.0 mL) was added oxalyl chloride (270 mg, 2.12mmol) followed by dimethylformamide (0.05 mL). The reaction mixture wasstirred under nitrogen atmosphere at room temperature for 2 h. Afterthis time, the reaction mixture was concentrated under reduced pressureto afford crude 4 (400 mg, IN-MRG-K-163-1) as an off white solid, whichwas used without further purification.

(S)-methyl 1-[4-(1H-pyrazol-1-yl)benzoyl]aziridine-2-carboxylate [5]

To a solution of 3 (600 mg, 3.03 mmol) in chloroform (10 mL) was added 4(756 mg, 3.63 mmol), followed by triethylamine (1.0 mL, 7.57 mmol) andthe reaction mixture was stirred under nitrogen atmosphere at roomtemperature for 16 h. After this time, the reaction mixture was dilutedwith water (10 mL). The layers were separated and the aqueous layer wasextracted with chloroform (2×20 mL). The combined organic layers werewashed with brine (20 mL), dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The resulting residue was purifiedby chromatography (silica gel, CH₃OH/CH₂Cl₂, gradient) to afford 5 [480mg, 60% for two steps] as an off white solid.

(S)-methyl2-[4-(1H-pyrazol-1-yl)benzamido]-3-{[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino}propanoate[7]

To a solution of 5 (450 mg, 1.66 mmol) in acetonitrile (10 mL) was added6 (776 mg, 4.15 mmol) followed by triethylamine (0.6 mL, 4.15 mmol). Thereaction mixture was stirred under nitrogen atmosphere at 80° C. for 16h. After this time, the reaction mixture was cooled and diluted withwater (10 mL). The layers were separated and the aqueous layer wasextracted with ethyl acetate (2×20 mL). The combined organic layers werewashed with brine (10 mL), dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The resulting residue was purifiedby MS-triggered preparative HPLC to afford 7.TFA (100 mg, 14%) as abrown red gum. MS-triggered Preparative HPLC purifications wereperformed using a Sunfire C18 column, OBD, 10μ (30×250 mm) with UVdetection at 220 nm using a solvent gradient program (15% to 55%acetonitrile/water with 0.1% trifluoroacetic acid).

(S)-methyl2-[4-(1H-pyrazol-1-yl)benzamido]-3-[(tertbutoxycarbonyl)[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]propanoate[8]

To a solution of 7 (100 mg, 0.23 mmol) in methylene chloride (5.0 mL)was added Boc anhydride (0.7 mL, 0.35 mmol) followed by triethylamine(0.06 mL, 0.47 mmol). The reaction mixture was stirred under nitrogenatmosphere at room temperature for 2 h. After this time, the reactionmixture was diluted with water (5.0 mL). The layers were separated andthe aqueous layer was extracted with chloroform (2×10 mL). The combinedorganic layers were washed with brine (5.0 mL), dried over anhydroussodium sulfate and concentrated under reduced pressure. The resultingresidue was purified by chromatography (silica gel, EtOAc/hexanes,gradient) to afford 8 (60 mg, 48%) as an off white solid.

(S)-2-[4-(1H-pyrazol-1-yl)benzamido]-3-[(tert-butoxycarbonyl)[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]propanoicacid [9]

To a solution of 8 (60 mg, 0.11 mmol) in a mixture of tetrahydrofuran(2.0 mL) and water (1.0 mL) was added lithium hydroxide monohydrate (8.9mg, 0.17 mmol). The reaction mixture was stirred at room temperature for2 h. After this time, the reaction mixture was diluted with water (5 mL)and acidified with 2 N aqueous HCl to pH 5. The mixture was extractedwith ethyl acetate (2×10 mL). The combined organic layers were washedwith brine (10 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford 9 (50 mg, 86%) as an off white solid.

tert-butyl[(S)-2-[4-(1H-pyrazol-1-yl)benzamido]-3-(4-methylpiperazin-1-yl)-3-oxopropyl][(1R,2S)-2-(4-fluorophenyl)cyclopropyl]carbamate[10]

To a solution of 9 (50 mg, 0.09 mmol) in tetrahydrofuran (3.0 mL) wasadded (3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one) (DEPBT,44 mg, 0.14 mmol) followed by imidazole (10 mg, 0.147 mmol). Thereaction mixture was stirred under nitrogen atmosphere at 0° C. for 40min. After this time, 1-methylpiperazine (15 mg, 0.14 mmol) was added at0° C. The reaction was warmed to room temperature and stirred undernitrogen atmosphere for 16 h. After this time, the reaction mixture wasfiltered through diatomaceous earth and rinsed with ethyl acetate (10mL). The filtrate was washed with brine (5.0 mL), dried over anhydroussodium sulfate and concentrated under reduced pressure. The resultingresidue was purified by chromatography (silica gel, CH₃OH/CH₂Cl₂,gradient) to afford 10 (30 mg, 51%) as an off white solid.

N—[(S)-3-{[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino}-1-(4-methylpiperazin-1-yl)-1-oxopropan-2-yl]-4-(1H-pyrazol-1-yl)benzamidehydrochloride salt

To a solution of 10 (30 mg, 0.05 mmol) in trifluoroethanol (3.0 mL) wasadded chlorotrimethylsilane (0.1 mL). The reaction mixture was stirredunder nitrogen atmosphere at 0° C. for 2 h. After this time, thereaction mixture was concentrated under reduced pressure. The resultingresidue was triturated with methyl tert-butyl ether and pentanes toafford the product (20 mg, 71%) as a hygroscopic off white solid.

Example 186

Synthesis of 186

Into a 250-mL round-bottom flask, was placed 4-(phenylsulfonyl)benzoicacid (2 g, 7.63 mmol, 1.00 equiv), sulfurooyl dichloride (20 mL). Theresulting solution was stirred for 16 h at 80° C. in an oil bath. Theresulting mixture was concentrated under vacuum. This resulted in 2 g(93%) of (1) as a off-white solid.

Into a 1000-mL round-bottom flask, was placed a solution of(2S)-2-amino-4-[(tert-butyldiphenylsilyl)oxy]butanoic acid (2.12 g, 5.93mmol, 1.00 equiv) in H₂O (50 ml), dioxane (50 mL), a solution of sodiumcarbonate (1.89 g, 17.79 mmol, 3 equiv) in water (20 mL). This wasfollowed by the addition of a solution of (1) (2 g, 7.12 mmol, 1.20equiv) in dioxane (40 mL) dropwise with stirring at 0° C. The resultingsolution was stirred for 1 h at 0° C. in a water/ice bath. The pH valueof the solution was adjusted to 2 with hydrogen chloride (2 mol/L). Theresulting solution was extracted with 3×100 mL of ethyl acetate and theorganic layers combined. The resulting mixture was washed with 1×100 mLof brine. The mixture was dried over anhydrous sodium sulfate. Theresulting mixture was concentrated under vacuum. The resulting mixturewas washed with 20 mL of DCM. The solids were filtered out. Thisresulted in 3 g (70%) of (2) as yellow oil.

Into a 100-mL 3-necked round-bottom flask, was placed a solution of (2)(3 g, 4.98 mmol, 1.00 equiv) in tetrahydrofuran (100 mL), DEPBT (2235mg, 7.48 mmol, 1.50 equiv). This was followed by the addition ofimidazole (508 mg, 7.48 mmol, 1.50 equiv), stirred for 40 mins at 0° C.To this was added a solution of Thiomorpholine-1,1-dioxide (807 mg, 5.98mmol, 1.20 equiv) in tetrahydrofuran (10 mL) dropwise with stirring at0° C. The resulting solution was stirred for 16 h at 25° C. The solidswere filtered out. The resulting mixture was concentrated under vacuum.The resulting solution was diluted with 100 mL of H₂O. The resultingsolution was extracted with 3×100 mL of ethyl acetate and the organiclayers combined and dried over anhydrous sodium sulfate. The residue wasapplied onto a silica gel column with ethyl acetate/petroleum ether(1:3). The collected fractions were combined and concentrated undervacuum. This resulted in 2.5 g (70%) of (3) as yellow oil.

Into a 50-mL round-bottom flask, was placed a solution of (3) (1500 mg,2.09 mmol, 1.00 equiv) in tetrahydrofuran (30 mL), TBAF (4.2 mL, 2.00equiv). The resulting solution was stirred for 16 h at 25° C. Theresulting mixture was concentrated under vacuum. The resulting solutionwas diluted with 50 mL of H₂O. The resulting solution was extracted with3×50 mL of ethyl acetate and the organic layers combined and dried overanhydrous sodium sulfate. The residue was applied onto a silica gelcolumn with ethyl acetate. The collected fractions were combined andconcentrated under vacuum. This resulted in 700 mg (70%) of (4) asyellow oil.

Into a 25-mL round-bottom flask, was placed a solution of (4) (200 mg,0.42 mmol, 1.00 equiv) in dichloromethane (20 mL), Dess-Martin (353 mg,0.83 mmol, 2.00 equiv). The resulting solution was stirred for 30 min at25° C. The residue was applied onto a silica gel column with ethylacetate. The collected fractions were combined and concentrated undervacuum. This resulted in 150 mg (75%) of (5) as a light yellow solid.

Into a 25-mL round-bottom flask, was placed a solution of (5) (150 mg,0.31 mmol, 1.00 equiv) in dichloromethane (20 mL),(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (57 mg, 0.38 mmol, 1.20equiv), NaBH(AcO)₃ (159 mg, 0.75 mmol, 2.40 equiv). The resultingsolution was stirred for 10 min at 25° C. The resulting solution wasdiluted with 30 mL of H₂O. The resulting solution was extracted with2×20 mL of dichloromethane and the organic layers combined and driedover anhydrous sodium sulfate and concentrated under vacuum. The crudeproduct was purified by Prep-HPLC. This resulted in 19.2 mg (10%) ofN—((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)-4-(phenylsulfonyl)benzamideas a white solid.

Example 187

Synthesis of 187

Into a 250-mL round-bottom flask, was placed a solution of(2S)-4-[(tert-butyldiphenylsilyl)oxy]-2-[[4-(4-fluorophenyl)phenyl]formamido]butanoicacid (2 g, 3.60 mmol, 1.00 equiv) in tetrahydrofuran (50 mL), DEPBT(1.62 g, 5.41 mmol, 1.50 equiv). This was followed by the addition ofimidazole (370 mg, 5.44 mmol, 1.50 equiv). stirred for 40 min at 0° C.To this was added a solution of thiomorpholine-1,1-dioxide (890 mg, 6.58mmol, 1.50 equiv) in tetrahydrofuran (20 mL) dropwise with stirring at0° C. in 30 min. The resulting solution was stirred for 16 h at 25° C.The resulting mixture was concentrated under vacuum. The resultingsolution was diluted with 50 mL of H₂O. The resulting solution wasextracted with 3×50 mL of ethyl acetate and the organic layers combined.The residue was applied onto a silica gel column with ethylacetate/petroleum ether (1:1). This resulted in 2 g (82%) of (1) as aoff-white solid.

Into a 100-mL round-bottom flask, was placed a solution ofN-[(2S)-4-[(tert-butyldiphenylsilyl)oxy]-1-oxo-1-(thiomorpholine-1,1-dioxide-4-yl)-butan-2-yl]-4-(4-fluorophenyl)-benzamide(2 g, 2.97 mmol, 1.00 equiv) in tetrahydrofuran (10 mL), TBAF (6 mL,2.00 equiv). The resulting solution was stirred for 16 h at 25° C. Theresulting mixture was concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate. This resulted in 1 g (77%)of (2) as a off-white solid.

Into a 50-mL round-bottom flask, was placed a solution of4-(4-fluorophenyl)-N-[(2S)-4-hydroxy-1-oxo-1-(thiomorpholine-1,1-dioxide-4-yl)-butan-2-yl]benzamide(200 mg, 0.46 mmol, 1.00 equiv) in dichloromethane (10 mL), Dess-Martin(389 mg, 0.92 mmol, 2.00 equiv). The resulting solution was stirred for1 h at 25° C. The residue was applied onto a silica gel column withethyl acetate. The collected fractions were combined and concentratedunder vacuum. This resulted in 150 mg (75%) of (3) as yellow oil.

Into a 50-mL round-bottom flask, was placed a solution ofN-[(2S)-1,4-dioxo-1-(thiomorpholine-1,1-dioxide-4-yl)-butan-2-yl]-4-(4-fluorophenyl)benzamide(150 mg, 0.35 mmol, 1.00 equiv) in dichloromethane (15 mL),(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (63.2 mg, 0.42 mmol, 1.20equiv), NaBH(AcO)₃ (147 mg, 0.69 mmol, 2.00 equiv). The resultingsolution was stirred for 10 min at 28° C. The resulting mixture wasconcentrated under vacuum. The crude product was purified by Prep-HPLC.This resulted in 37.1 mg (19%) of4′-fluoro-N—((S)-4-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-oxo-1-(thiomorpholino-4,4-dioxide-1-yl)-butan-2-yl)biphenyl-4-carboxamideas a white solid.

Example 188

Synthesis of 188

Into a 500-mL round-bottom flask, was placed a solution of(2S)-2-[(4-fluorophenyl)formamido]-5-methoxy-5-oxopentanoic acid (3.4 g,12.00 mmol, 1.00 equiv) in tetrahydrofuran (200 mL), DEPBT (5.4 g, 1.50equiv). This was followed by the addition of imidazole (1.2 g, 1.50equiv). stirred for 40 min at 0° C. To this was added azetidine (1.1 g,19.27 mmol, 1.50 equiv) dropwise with stirring at 0° C. in 10 min. Theresulting solution was stirred for 16 h at 20° C. The resulting mixturewas concentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (10:1). The collectedfractions were combined and concentrated under vacuum. This resulted in3.5 g (90%) of PH-(1) as yellow oil.

Into a 250-mL round-bottom flask, was placed a solution of methyl(4S)-5-(azetidin-1-yl)-4-[(4-fluorophenyl)formamido]-5-oxopentanoate (2g, 6.20 mmol, 1.00 equiv) in tetrahydrofuran (100 mL), a solution oflithiumol (270 mg, 11.27 mmol, 1.80 equiv) in water (20 mL). Theresulting solution was stirred for 2 h at 20° C. The resulting mixturewas concentrated under vacuum. This resulted in 1.5 g (78%) of (2) as alight yellow solid.

Into a 100-mL round-bottom flask, was placed a solution of(4S)-5-(azetidin-1-yl)-4-[(4-fluorophenyl)formamido]-5-oxopentanoic acid(500 mg, 1.62 mmol, 1.00 equiv) in tetrahydrofuran (20 mL). This wasfollowed by the addition of NMM (350 mg, 3.46 mmol, 2.10 equiv). stirredfor 40 min at 0 degrees. To this was addedchloro(2-methylpropoxy)methanone (470 mg, 3.44 mmol, 2.10 equiv).stirred for 60 min at −20° C. To the mixture was added NaBH₄ (650 mg,17.18 mmol, 10.00 equiv), methanol (5 mL). The resulting solution wasstirred for 1 h at 30° C. in an oil bath. The residue was applied onto asilica gel column with ethyl acetate/petroleum ether (10:1). Thecollected fractions were combined and concentrated under vacuum. Thisresulted in 350 mg (73%) of (3) as yellow oil.

Into a 100-mL round-bottom flask, was placed a solution ofN-[(2S)-1-(azetidin-1-yl)-5-hydroxy-1-oxopentan-2-yl]-4-fluorobenzamide(350 mg, 1.19 mmol, 1.00 equiv) in dichloromethane (20 mL), Dess-Martin(1 g, 2.36 mmol, 2.00 equiv). The resulting solution was stirred for 60min at 30° C. in an oil bath. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (10:1). The collectedfractions were combined and concentrated under vacuum. This resulted in230 mg (66%) of (4) as yellow oil.

Into a 100-mL round-bottom flask, was placed a solution ofN-[(2S)-1-(azetidin-1-yl)-1,5-dioxopentan-2-yl]-4-fluorobenzamide (230mg, 0.79 mmol, 1.00 equiv) in dichloromethane (20 mL),(1R,2S)-2-(4-fluorophenyl)cyclopropan-1-amine (176 mg, 1.16 mmol, 1.20equiv), NaBH(AcO)₃ (422 mg, 2.40 equiv). The resulting solution wasstirred for 5 min at 20° C. The resulting mixture was concentrated undervacuum. The crude product (1 mL) was purified by Flash-Prep-HPLC. Thisresulted in 86 mg (26%) ofN—((S)-1-(azetidin-1-yl)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-oxopentan-2-yl)-4-fluorobenzamideas a off-white solid.

The following table discloses physical data for compounds; examples notlisted above were synthesized according to the methods disclosed above,form example according to the identified scheme. Such compounds may alsobe synthesized by those of skill in the art according to methods knownin the art.

TABLE 2 Physical Data and Synthetic Methods Ex # Scheme MS 1H NMR 1 1492 [M + H]⁺ (300 MHz, CD₃OD-d₄) δ ppm: 8.61 (d, J = 1.3 Hz, 1H), 8.06(d, J = 8.6 Hz, 2H), 8.02-7.94 (m, 2H), 7.90 (d, J = 1.3 Hz, 1H), 7.06(ddt, J = 8.0, 5.3, 2.7 Hz, 2H), 6.93 (t, J = 8.8 Hz, 2H), 5.33-5.17 (m,1H), 3.78-3.41 (m, 4H), 3.07 (dd, J = 6.9, 5.4 Hz, 2H), 2.55-2.28 (m,5H), 2.26 (s, 3H), 1.90 (ddt, J = 9.1, 5.9, 2.8 Hz, 1H), 1.15- 0.87 (m,2H). 2 1 503 (M + 1) (400 MHz, Methanol-d₄) δ 8.88 (d, J = 4.9 Hz, 2H),8.55- 8.47 (m, 2H), 8.04-7.96 (m, 2H), 7.40 (t, J = 4.9 Hz, 1H), 7.10(ddd, J = 8.2, 5.2, 2.5 Hz, 2H), 6.96 (td, J = 8.8, 1.6 Hz, 2H),5.34-5.23 (m, 1H), 3.71 (s, 2H), 3.64 (d, J = 16.6 Hz, 1H), 3.59-3.51(m, 3H), 3.18-3.02 (m, 2H), 2.54-2.31 (m, 5H), 2.29 (s, 3H), 1.93 (ddt,J = 9.4, 6.2, 3.4 Hz, 1H), 1.16-0.95 (m, 2H). 3 1 450 [M + H]⁺ (300 MHz,CD₃OD-d₄) δ ppm: 8.03-7.92 (m, 2H), 7.87- 7.76 (m, 2H), 7.05 (ddd, J =8.4, 5.3, 2.6 Hz, 2H), 6.98- 6.86 (m, 2H), 5.30-5.11 (m, 1H), 3.73-3.42(m, 4H), 3.05 (dd, J = 7.2, 5.4 Hz, 2H), 2.49-2.28 (m, 5H), 2.25 (s,3H), 1.89 (dt, J = 9.3, 4.4 Hz, 1H), 1.15-0.88 (m, 2H). 4 2 550 [M + H]⁺(300 MHz, MeOD-d₄) δ ppm: 8.01(m, 2H), 7.81-7.77(m, 2H), 7.71-7.66(m,2H), 7.50-7.45(m, 2H), 7.42-7.38(m, 1H), 7.22-7.17(m, 2H), 7.10-7.01(m,2H), 5.28-5.20(m, 1H), 4.39-4.31(m, 1H), 4.19-4.10(m, 1H), 3.78-3.98(m,2H), 3.36-3.32(m, 2H), 3.21-3.11(m, 4H), 3.05-2.98(m, 1H), 2.57-2.51(m,1H), 2.39-2.22(m, 1H), 2.21-2.11(m, 1H), 1.57-1.49(m, 1H), 1.42-1.37(m,1H) 5 2 515 [M + H]⁺ (300 MHz, Methanol-d₄) δ 7.97-7.88 (m, 2H),7.72-7.63 (m, 4H), 7.47-7.33 (m, 4H), 7.05-7.00(m, 2H), 6.93-6.88 (m,2H), 5.14-5.10 (m, 1H), 3.75-3.50 (m, 4H), 2.84-2.79 (m, 2H), 2.44-2.40(m, 4H), 2.32-2.25 (m, 4H), 2.09-1.85 (m, 3H), 1.05-1.00 (m, 1H),0.97-0.93 (m, 1H) 6 2 [M + H]+472 (300 MHz, MeOD): δ d7.88~7.90(d, J =6.0 Hz, 2H), 7.62~7.71(m, 4H), 7.41~7.46(t, J = 7.35 Hz, 2H),7.35~7.37(m, 1H), 7.00~7.05(m, 2H), 6.88~6.93(t, J = 8.7 Hz, 2H),4.61~4.66(m, 1H), 4.47~4.49(m, 1H), 4.29~4.31(m, 1H), 4.00~4.05(m, 2H),2.81~2.85(t, J = 6.9 Hz, 2H), 2.26~2.34(m, 3H), 1.88~2.01(m, 3H),0.93~1.06(m, 2H). 7 2 597 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm:8.00-7.92(m, 2H), 7.79- 7.68(m, 4H), 7.29-7.20 (m, 4H), 7.07-6.98 (m,2H), 5.23- 5.19 (m, 1H), 3.98-3.87 (m, 1H), 3.86-3.72(m, 1H), 3.69- 3.59(m, 2H), 3.37 (m, 4H), 3.26-3.10 (m, 2H), 3.02-2.95 (m, 1H), 2.82(s,3H), 2.57-2.49 (m, 1H), 2.39-2.25 (m, 1H), 2.21-2.09 (m, 1H),1.55-1.47(m, 1H), 1.42-1.35(m, 1H) 8 2 550 [M + H]⁺ (300 MHz,Methanol-d₄) δ 8.15-8.14 (m, 1H), 7.89- .786 (m, 2H), 7.70-7.68 (m, 2H),7.63-7.59 (m, 1H), 7.51-7.47 (m, 2H), 7.42-7.41 (m, 2H), 7.23-7.20 (m,2H), 7.06-7.02 (m, 2H), 5.28-5.24 (m, 1H), 4.39-4.34 (m, 1H), 4.21-4.13(m, 1H), 3.96-3.81 (m, 2H), 3.37-3.32 (m, 2H), 3.19-3.16 (m, 4H),3.02-3.00 (m, 1H), 2.52-2.49 (m, 1H), 2.38-2.31 (m, 1H), 2.26-2.18 (m,1H), 1.55-1.52 (m, 1H), 1.41-1.39 (m, 1H) 9 2 [M + H]+540 (300 MHz,DMSO): 8.44 (d, J = 8 Hz, 2H), 7.70-7.51 (m, 4H), 7.50-7.40 (m, 2H),7.40-7.28 (m, 3H), 7.08-6.96 (m, 4H), 4.90-4.70 (m, 1H), 3.75-3.45 (m,6H), 3.20-2.95 (m, 4H), 2.88 (s, 3H), 2.60-2.55 (m, 2H), 2.20~2.05 (m,1H), 1.82-1.65 (m, 3H), 0.95-0.70 (m, 2H) 10 2 567 [M + H]⁺ (300 MHz,Methanol-d₄) δ 7.78-7.75 (m, 4H), 7.44- 7.40 (m, 3H), 6.95-6.85 (m, 4H),4.92-4.81(m, 1H), 3.90- 3.68 (m, 4H), 3.54-3.36 (m, 2H), 3.23-3.11 (m,2H), 3.03-2.86 (m, 2H), 2.68-2.63 (m, 5H), 2.11-2.07 (m, 1H), 1.88-1.73(m, 3H), 0.93-0.88 (m, 1H), 0.86-0.79 (m, 1H) 11 2 479 [M + H]⁺ (300MHz, MeOD-d₄) δ ppm: 9.34 (s, 1H), 9.00 (s, 1H), 8.23-8.10 (m, 2H),8.05-7.90 (m, 1H), 7.90-7.72 (m, 1H), 7.30-7.12 (m, 2H), 7.12-6.93 (m,2H), 5.40-5.18 (m, 1H), 4.00-3.80 (m, 2H), 3.80-3.60 (m, 2H), 3.45-3.35(m, 4H), 3.30-3.18 (m, 2H), 3.10-2.98 (m, 1H), 2.86 (s, 3H), 2.62- 2.50(m, 1H), 2.58-2.15 (m, 2H), 1.65-1.50 (m, 1H), 150- 1.31(m, 1H) 12 3[M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.31 (s, 1H), 8.05-7.84 540 (m, 4H),7.75 (d, J = 1.7 Hz, 1H), 7.23-7.12 (m, 2H), 7.08-6.94 (m, 2H),6.50-6.62 (m, 1H), 5.20-5.30 (m, 1H), 4.42-4.25 (m, 1H), 4.20-4.01 (m,1H), 3.98-3.72 (m, 2H), 3.29-3.35 (m, 2H), 3.10-3.20 (m, 4H), 3.05-2.97(m, 1H), 2.58-2.42 (m, 1H), 2.39-2.05 (m, 2H), 1.58-1.42 (m, 1H),1.42-1.30 (m, 1H). 13 2 548[M + H]⁺ (300 MHz, CD₃OD-d₄) δ ppm: 7.91-7.82(m, 2H), 7.45-7.40 (m, 2H), 7.23-7.15 (m, , 1H), 7.10-7.00 (m, 6H),7.00-6.88 (m, 2H), 5.15-5.10 (m, 1H), 4.55- 4.28 (m, 2H), 4.01-3.85 (m,1H), 3.75-3.60 (m, 1H), 3.45- 3.35(m, 1H), 3.25-3.05 (m, 3H), 3.95-3.75(m, 2H), 3.35- 3.21 (m, 1H), 2.11-1.85 (m, 3H), 1.12-0.85 (m, 2H) 14 2553 [M + H]⁺ (400 MHz, Methanol-d₄) δ 8.47-8.23 (m, 1H), 8.08- 7.52 (m,2H), 7.75-7.68 (m, 1H), 7.63-7.49(m, 3H), 7.08- 6.91 (m, 4H), 5.31-5.29(M, 1H), 4.13-3.89 (m, 2H), 3.87-3.71 (m, 1H), 3.64-3.47 (m, 2H),3.26-3.16 (m, 1H), 2.95-2.80 (m, 6H), 2.37-2.31 (m, 1H), 2.24-.86 (m,4H), 1.12-1.01(m, 1H), 1.00-0.88 (m, 1H) 15 2 583 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 8.44 (s, 1H), 8.22-7.71 (m, 4H), 7.73-7.42 (m, 2H), 7.25(dd, J = 8.3, 5.1 Hz, 1H), 7.20-6.77 (m, 3H), 4.77 (t, J = 9.4 Hz, 1H),3.95 (s, 1H), 3.63-3.47 (m, 2H), 3.25-2.70 (m, 3H), 2.64- 2.44 (m, 1H),2.35 (dq, J = 9.2, 4.8, 4.2 Hz, 1H), 2.24- 1.85 (m, 2H), 1.62-0.90 (m,2H). 16 2 [M + H]+540 (300 MHz, MeOD): 8.10~7.95 (m, 2H), 7.92-7.75 (m,2H), 7.10-6.90 (m, 4H), 5.20~5.10 (m, 1H), 4.08~3.88 (m, 2H), 3.85-3.68(m, 1H), 3.65~3.50 (m, 2H), 3.26~3.10 (m, 1H), 2.95~2.81 (m, 5H),2.40~2.30(m, 1H), 2.20~1.90(m, 3H), 1.15~1.00 (m, 1H), 1.00~0.92 (m,1H). 17 2 579 [M + H]⁺ (300 MHz, Methanol-d₄) δ 8.11-8.10 (m, 1H), 7.84-.781 (m, 2H), 7.65-7.62 (m, 2H), 7.58-7.52 (m, 1H), 7.46-7.41 (m, 2H),7.37-7.33 (m, 1H), 7.18-7.13 (m, 2H), 7.01-6.95 (m, 2H), 5.20-5.16 (m,1H), 3.91-3.73 (m, 2H), 3.63-3.55 (m, 2H), 3.41-3.29 (m, 3H), 3.26-3.21(m, 1H), 3.20-3.09 (m, 2H), 2.98-2.94 (m, 1H), 2.78 (s, 3H), 2.53-2.44(m, 1H), 2.37-2.10 (m, 2H), 1.51-1.48 (m, 1H), 1.37-1.33(m, 1H) 18 2591[M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 7.98 (dd, J = 8.2, 1.4 Hz, 2H),7.78 (d, J = 8.0 Hz, 2H), 7.69 (d, J = 7.5 Hz, 2H), 7.49 (t, J = 7.5 Hz,2H), 7.45-7.38 (m, 1H), 7.12 (d, J = 8.2 Hz, 2H), 6.87 (d, J = 8.4 Hz,2H), 5.22 (dd, J = 8.1, 5.4 Hz, 1H), 4.93 (s, 1H), 3.90 (d, J = 13.3 Hz,1H), 3.77 (d, J = 1.3 Hz, 4H), 3.65 (dd, J = 13.1, 6.6 Hz, 2H), 3.36 (s,3H), 3.19 (q, J = 10.0 Hz, 2H), 2.95 (dt, J = 7.9, 4.0 Hz, 1H), 2.85 (s,3H), 2.48 (ddd, J = 10.3, 6.6, 3.5 Hz, 1H), 2.33 (dq, J = 13.8, 7.3 Hz,1H), 2.20 (dq, J = 14.4, 7.3 Hz, 1H), 1.50 (ddd, J = 10.7, 6.7, 4.4 Hz,1H), 1.35 (q, J = 7.1 Hz, 1H). 19 2 562 [M + H]⁺ (300 MHz, Methanol-d4)δ 7.9-7.88 (m, 2H), 7.71-764 (m, 4H), 7.46-7.43 (m, 2H), 7.41-7.35 (m,1H), 6.95- 6.92 (m, 2H), 6.76-6.73 (m, 2H), 5.14-5.09 (m, 1H), 4.45-4.27(m, 2H), 3.93-3.84 (m, 1H), 3.72-3.61 (m, 4H), 3.45-3.33 (m, 1H),3.19-3.04 (m, 3H), 2.88-2.83 (m, 2H), 2.25-2.22 (m, 1H), 2.02-1.96 (m,2H), 1.84-1.81 (m, 1H), 1.00-0.93 (m, 1H), 0.91-0.88 (m, 1H) 20 4 657[M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.09 (d, J = 8.1 Hz, 2H), 8.06-8.02(m, 2H), 7.97 (d, J = 8.2 Hz, 2H), 7.88 (d, J = 8.0 Hz, 2H), 7.23 (dd, J= 8.5, 5.4 Hz, 2H), 7.06 (t, J = 8.7 Hz, 2H), 5.24 (dd, J = 8.0, 5.5 Hz,1H), 3.90 (s, 1H), 3.79 (s, 1H), 3.67 (d, J = 12.2 Hz, 2H), 3.36 (d, J =11.8 Hz, 4H), 3.20 (s, 5H), 3.02 (dt, J = 7.9, 4.0 Hz, 1H), 2.86 (s,3H), 2.55 (d, J = 9.1 Hz, 1H), 2.41-2.27 (m, 1H), 2.27-2.12 (m, 1H),1.56 (dt, J = 10.8, 5.5 Hz, 1H), 1.41 (q, J = 7.2 Hz, 1H). 21 5 541 [M +H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.25-8.15 (m, 2H), 8.15- 8.00 (m, 3H),7.42-7.37 (m, 1H), 7.28-7.15 (m, 2H), 7.10-7.00 (m, 2H), 5.30-5.20 (m,1H), 4.48-4.29 (m, 1H), 4.25-4.10 (m, 1H), 4.00-3.85 (m, 2H), 3.42-3.37(m, 2H), 3.27-3.12 (m, 46H), 3.12-2.92 (m, 1H), 2.60-2.50 (m, 1H),2.45-2.30 (m, 1H), 2.30-2.17 (m, 1H), 1.60-1.50 (m, 1H), 1.50-1.40 (m,1H) 22 4 [M + H]+ (300 MHz, MeOD-d₄): δ ppm 8.10-7.90 (m, 2H), 7.82- 5557.70 (m, 2H), 7.22-7.15 (m, , 2H), 7.05-6.85 (m, 2H), 5.30-5.15 (m, 1H),4.55-4.25 (m, 1H), 4.25-4.03 (m, 1H), 4.05-3.70 (m, 2H), 3.35-3.30 (m,2H), 3.25-3.05 (m, 4H), 3.05-2.88 (m, 1H), 2.60-2.45 (m, 1H), 2.45-2.05(m, 2H), 1.70-1.50 (m, 1H), 1.45-1.29 (m, 1H). 23 6 506 [M + H]⁺ (400MHz, MeOD-d₄) δ ppm: 8.64 (d, J = 1.2 Hz, 1H), 8.10-7.96 (m, 4H), 7.93(d, J = 1.2 Hz, 1H), 7.04 (m, 2H), 6.98-6.87 (m, 2H), 5.06 (m, 1H), 3.68(m, 3H), 3.55-3.44 (m, 1H), 2.96-2.70 (m, 6H), 2.28 (m, 1H), 1.95-1.75(m, 3H), 1.66 (m, 2H), 1.09-0.91 (m, 2H). 24 6 [M + H]⁺477 (300 MHz,CD₃OD)δ ppm: 8.62 (s, 1H), 7.96-8.15(m, 4H), 7.85-7.96 (m, 1H),7.08-7.24 (m, 2H), 6.90-7.08 (m, 2H), 4.56-4.75 (m, 1H), 4.39-4.52 (m,1H), 4.18-4.38(m, 1H), 3.86-4.12 (m, 2H), 3.18-3.25 (m, 2H), 2.90-3.00(m, 1H), 2.38-2.40(m, 1H), 2.23-2.40 (m, 2H), 1.70-2.00 (m, 4H),1.40-1.54 (m, 1H), 1.20-1.40 (m, 1H). 25 6 534 [M + H] (400 MHz,MeOD-d₄) δ ppm: 8.64(s, 1H), 8.09-8.01 m, 4H), 7.94(s, 1H), 7.20-7.17(m,2H), 7.04-6.99(m, 2H), 4.90-4.87(m, 1H), 4.16-3.87(m, 3H), 3.82-3.69(m,1H), 3.1-3.42(m, 1H), 3.27-3.22(m, 2H), 2.99-2.94(m, 7H), 2.58-2.43(m,2H), 2.37-2.15(m, 1H), 2.01-1.82(m, 4H), 1.50-1.47(m, 1H), 1.39-1.35(m,1H) 26 6 548 [M + H] (300 MHz, MeOD-d₄) δ ppm: 8.61 (s, 1H), 8.10-7.88(m, 5H), 7.15 (m, 2H), 6.97 (m, 2H), 5.09 (s, 1H), 4.75- 4.63 (m, 1H),4.29 (m, 1H), 3.47 (m, 1H), 3.21 (m, 4H), 2.98-2.85 (m, 6H), 2.71 (m,1H), 2.47 (m, 1H), 2.13 (m, 2H), 1.93-1.72 (m, 5H), 1.61 (m, 1H), 1.48(m, 1H), 1.31 (m, 1H). 27 6 520 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.21(s, 2H), 8.10 (d, J = 8.3 Hz, 2H), 7.90-7.78 (m, 2H), 7.28-7.16 (m, 2H),7.09-6.98 (m, 2H), 5.12 (dd, J = 8.2, 5.2 Hz, 1H), 4.60 (d, J = 78.2 Hz,2H), 3.59 (s, 3H), 3.31-3.04 (m, 4H), 3.03-2.87 (m, 4H), 2.50 (ddd, J =10.3, 6.6, 3.5 Hz, 1H), 1.95 (dddd, J = 28.6, 20.2, 15.7, 8.6 Hz, 4H),1.51 (ddd, J = 10.7, 6.9, 4.4 Hz, 1H), 1.38 (q, J = 7.1 Hz, 1H). 28 6[M + H]⁺ (300 MHz, MeOD-d₄): δ ppm 8.85 (d, J = 4.9 Hz, 2H), 5618.52-8.42 (m, 2H), 7.98-7.89 (m, 2H), 7.37 (t, J = 4.9 Hz, 1H),7.07-6.83 (m, 4H), 5.08-5.04 (m, 1H), 3.76- 3.51 (m, 6H), 2.78-2.72 (m,2H), 2.60-2.50 (m, 6H), 2.30- 2.19 (m, 1H), 1.93-1.74 (m, 3H), 1.68-1.62(m, 2H), 1.07-0.86 (m, 2H) 29 6 435 [M + H] (400 MHz, MeOD-d₄) δ ppm:8.31 (d, J = 2.6 Hz, 1H), 8.00-7.81 (m, 4H), 7.75 (d, J = 1.7 Hz, 1H),7.22-7.11 (m, 2H), 7.07-6.94 (m, 2H), 6.55 (dd, J = 2.6, 1.8 Hz, 1H),4.65 (t, J = 6.8 Hz, 1H), 4.35 (dq, J = 39.0, 8.3 Hz, 2H), 4.13-3.91 (m,2H), 3.22 (t, J = 7.3 Hz, 2H), 2.93 (dt, J = 8.0, 4.1 Hz, 1H), 2.49-2.23(m, 3H), 1.84 (q, J = 11.1, 8.9 Hz, 4H), 1.51-1.27 (m, 2H) 30 6 524 [M +H] (400 MHz, MeOD-d₄) δ ppm: 7.96-7.94(d, J = 8 Hz, 2H), 7.83-7.81(d, J= 8 Hz, 2H), 7.05-7.02(m, 2H), 6.94- 6.90(m, 2H), 5.10-5.07(m, 1H),4.51-4.42(m, 1H), 4.17- 4.10(m, 1H), 3.51-3.40(m, 1H), 3.09-2.95(m, 1H),2.78- 2.72(m, 2H), 2.30-1.75(m, 8H), 1.69-1.57(m, 2H), 1.05- 0.92(m,2H), 31 6 550 [M + H] (400 MHz, MeOD-d₄) δ ppm: 8.02-7.95 (d, J = 8.4Hz, 2H), 7.89-7.81 (d, J = 8.4 Hz, 2H), 7.23-7.13 (m, 2H), 7.07-6.97 (m,2H), 5.09 (m, 1H), 4.29 (m, 2H), 4.03 (m, 4H), 3.87 (s, 1H), 3.71 (s,1H), 3.50-3.17 (m, 6H), 2.93 (m, 1H), 2.46 (m, 1H), 2.00-1.80 (m, 4H),1.48 (m, 1H), 1.34 (m, 4H) 32 6 542[M + H]⁺ (300 MHz, Methanol-d4) δ8.33 (d, J = 4.8 Hz, 2H), 8.00- 7.91 (m, 2H), 7.87-7.77 (m, 2H), 7.15(ddd, J = 8.5, 5.2, 2.6 Hz, 2H), 7.06-6.91 (m, 2H), 6.62 (t, J = 4.8 Hz,1H), 5.13 (dd, J = 7.7, 5.3 Hz, 1H), 3.75 (m, 8H), 3.28- 3.15 (m, 2H),2.92 (dt, J = 7.9, 4.1 Hz, 1H), 2.41 (ddd, J = 10.4, 6.6, 3.6 Hz, 1H),2.05-1.74 (m, 4H), 1.50-1.27 (m, 2H) 33 6 577 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 8.88-8.87(m, 2H), 8.51- 8.49(m, 2H), 7.97-7.95(m, 2H),7.41-7.38(m, 1H), 7.05- 7.02(m, 2H), 6.93-6.89(m, 2H), 5.13-5.10(m, 1H),4.53- 4.42(m, 1H), 4.19-4.12(m, 1H), 3.53-3.41(m, 1H), 3.12- 2.95(m,1H), 2.78-2.72(m, 2H), 2.30-1.76(m, 8H), 1.71- 1.61(m, 2H), 1.05-1.01(m,1H), 0.97-0.92(m, 1H) 34 6 530 [M + H]⁺ ¹H NMR (300 MHz, Methanol-d₄) δ8.80 (d, J = 4.8 Hz, 2H), 8.50 (d, J = 8.4 Hz, 2H), 7.98 (d, J = 8.1 Hz,2H), 7.41 (t, J = 4.8 Hz, 1H), 7.11-6.86 (m, 4H), 4.85-4.80 (m, 4H),4.75-4.62 (m, 1H), 4.60-4.44 (m, 2H), 4.28-4.10 (m, 2H), 2.81-2.75 (m,2H), 2.40-2.20 (m, 1H), 1.95-1.83 (m, 3H), 1.75-1.60 (m, 2H), 1.10-0.95(m, 3H). 35 7 501[M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.42-8.22(d, J = 2.1Hz, 1H), 8.11-7.91 (m, 2H), 7.91-7.81 (m, 2H), 7.81-7.69 (m, 1H),7.18-7.08 (m, 2H), 7.08-6.88 (m, 2H), 6.81-6.52 (m, 1H), 4.85-4.45 (m,3H), 4.45-4.05 (m, 2H), 3.96-3.61 (m, 1H), 2.91-2.78 (m, 2H), 2.41-2.21(m, 1H), 1.95-1.71 (m, 3H), 1.46-1.71 (m, 2H), 1.15-0.85 (m, 2H), 36 7533 (M + 1) (CD₃OD, ppm): 8.36(s, 1H), 8.07-7.95 (m, 2H), 7.95- 7.87 (m,2H), 7.87-7.75 (m, 1H), 7.30-7.15 (m, 2H), 7.12-7.00 (m, 2H), 6.59 (s,1H), 5.05-5.20 (m, 1H), 4.20- 4.50 (m, 2H), 3.72-4.20 (m, 3H), 3.41-3.58(m, 2H), 3.15-3.28 (m, 2H), 2.90-3.05 (m, 4H), 2.43-2.53 (m, 1H),1.80-2.10 (m, 1H), 1.45-1.52 (m, 1H), 1.35-1.44 (m, 1H) 37 7 565 [M +H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.32(s, 1H), 7.99-7.97(d, J = 14.4 Hz,2H), 7.87-7.85(d, 2H), 7.76(s, 1H), 7.06- 7.02(m, 2H), 6.94-6.89(m, 2H),6.57-6.55(m, 1H), 5.12- 5.08(m, 1H), 4.19-4.12(m, 1H), 3.57-3.41(m, 1H),3.12- 2.95(m, 1H), 2.80-2.72(m, 2H), 2.29-1.76(m, 8H), 1.71- 1.61(m,2H), 1.06-1.01(m, 1H), 0.97-0.93(m, 1H), 38 7 518 [M + H]⁺ (300 MHz,Methanol-d₄) δ 8.34(s, 1H), 8.05-7.70(m, 5H), 7.10-6.90 (m, 4H), 6.57(d, J = 1.8 Hz, 1H), 4.87- 4.80 (m, 3H), 4.70-4.60 (m, 1H), 4.60-4.55(m, 2H), 4.28-4.20 (m, 1H), 4.20-4.08 (m, 1H), 2.82-2.72 (m, 2H),2.35-2.25 (m, 1H), 2.00-1.75 (m, 3H), 1.75-1.60 (m, 2H), 1.35-1.25 (m,1H), 1.10-1.00 (m, 3H). 41 7 548 (M + 1) (CD₃OD, ppm): 8.64 (s, 1H),7.88-8.15 (m, 5H), 7.01- 7.12 (m, 2H), 6.85-6.99 (m, 2H), 5.05-5.18 (m,1H), 4.28-4.45 (m, 1H), 3.95-4.11 (m, 1H), 2.95-3.10 (m, 1H), 2.71-2.85(m, 2H), 2.42-2.65 (m, 1H), 2.11-2.38 (m, 6H), 1.80-1.98 (m, 3H),1.55-1.71 (m, 2H), 1.27-1.36 (m, 2H), 1.12-1.22 (m, 6H), 0.85-1.10 (m,3H). 42 7 516 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.63(s, 1H), 8.08-8.04(m, 2H), 8.00-7.93(m, 2H), 7.93(s, 1H), 7.07-7.02(m, 2H), 6.94-6.90(m,2H), 4.83-4.72(m, 2H), 4.02-3.50(m, 2H), 2.82-2.77(m, 2H), 2.41-2.07(m,5H), 2.02-1.60(m, 5H), 1.06-1.03(m, 1H), 0.97-0.95(m, 1H) 43 7 [M + H]⁺541 (400 MHz, Methanol-d₄) δ 8.64 (s, 1H), 8.10-8.02 (m, 2H), 8.02-7.98(m, 2H), 7.97-7.91 (m, 1H), 7.11-7.03 (m, 2H), 7.00-6.88 (m, 2H),5.14-5.06 (m, 1H), 4.10-3.90 (m, 2H), 3.78-3.62 (m, 1H), 3.58-3.42 (m,1H), 2.86-2.70 (m, 2H), 2.36-2.26 (m, 1H), 2.25-1.80 (m, 7H), 1.80-1.61(m, 2H), 1.15-1.03 (m, 1H), 1.00-0.89 (m, 1H). 44 7 530 [M + H]⁺ (300MHz, MeOD-d₄) δ ppm: 8.60 (m, 1H), 8.11-7.93 (m, 4H), 7.90 (m, 1H),7.13-6.72 (m, 4H), 5.04 (m, 1H), 4.11-3.75 (m, 4H), 3.15-2.75 (m, 1H),2.75 (m, 2H), 2.25 (m, 1H), 2.18-1.75 (m, 7H), 1.64 (m, 2H), 1.05-0.97(m, 2H). 45 7 583 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61 (s, 1H),8.11-7.93 (m, 4H), 7.90 (d, J = 1.2 Hz, 1H), 7.02 (m, 2H), 6.90 (m, 2H),5.07 (m, 1H), 4.64 (m, 1H), 4.32 (m, 1H), 3.40-3.38 (m, 1H), 3.20-3.15(m, 1H), 2.91-2.76 (m, 6H), 2.28- 2.05(m, 3H), 2.05-1.76 (m, 7H),1.20-0.78 (m, 2H). 46 7 534 (M + 1) (CD₃OD, ppm): 8.64 (d, J = 1.2 Hz,1H), 8.10-8.05 (m, 2H), 8.05-8.00 (m, 2H), 7..98-7.87 (m, 1H), 7.10-7.00(m, 2H), 6.95-6.86 (m, 2H), 5.15-5.00 (m, 1H), 3.90-3.62 (m, 3H),3.60-3.50 (m, 1H), 2.85-2.70 (m, 2H), 2.65-2.40 (m, 6H), 2.35-2.27 (m,1H), 2.00-1.60 (m, 5H), 1.35-0.85 (m, 7H) 47 7 548 (M + 1) (CD₃OD, ppm):8.64 (s, 1H), 8.12-8.05 (m, 2H), 8.05- 8.00 (m, 2H), 8.00-7.95 (m, 1H),7.10-7.02 (m, 2H), 7.00-6.85 (m, 2H), 5.14-5.00 (m, 1H), 3.86-3.60 (m,3H), 3.60-3.50 (m, 1H), 2.90-2.70 (m, 3H), 2.70-2.0 (m, 4H), 2.35-2.20(m, 1H), 1.98-1.76 (m, 3H), 1.75-1.55 (m, 2H), 1.20-1.00 (m, 7H),1.00-0.90 (m, 1H) 48 7 584 (M + 1) (CD₃OD, ppm): 8.63 (s, 1H), 7.88-8.11(m, 5H), 7.00- 7.10 (m, 2H), 6.88-6.98 (m, 2H), 5.00-5.14 (m, 1H),3.85-4.02 (m, 2H), 3.63-3.78 (m, 1H), 3.45-3.60 (m, 1H), 3.37-3.45 (m,1H), 3.10-3.20 (m, 1H), 2.98-3.05 (m, 2H), 2.69-2.92 (m, 5H), 2.22-2.38(m, 1H), 1.79-1.98 (m, 3H), 1.60-1.79 (m, 2H), 0.91-1.10 (m, 2H). 49 7550 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.62 (d, J = 1.2 Hz, 1H),8.10-7.88 (m, 5H), 7.22-7.10 (m, 2H), 6.99 (m, 2H), 5.10 (m, 1H), 3.75(m, 6H), 3.21 (m, 2H), 3.10-2.89 (m, 7H), 2.43 (m, 1H), 1.99-1.80 (m,4H), 1.51-1.23 (m, 2H) 50 7 564 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61(s, 1H), 8.15-7.95 (m, 4H), 7.91 (d, J = 1.2 Hz, 1H), 7.15 (m, 2H),7.07- 6.85 (m, 2H), 5.08 (m, 1H), 4.30-3.78 (m, 3H), 3.71 (m, 2H), 3.38(m, 9H), 3.25 (m, 3H), 2.91 (m, 1H), 2.45 (m, 1H), 2.08-1.63 (m, 4H),1.51-1.18 (m, 2H). 51 7 520 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.31 (d,J = 2.6 Hz, 1H), 8.00-7.81 (m, 4H), 7.75 (d, J = 1.7 Hz, 1H), 7.22-7.11(m, 2H), 7.07-6.94 (m, 2H), 6.55 (dd, J = 2.6, 1.8 Hz, 1H), 4.65 (t, J =6.8 Hz, 1H), 4.35 (dq, J = 39.0, 8.3 Hz, 2H), 4.13-3.91 (m, 2H), 3.22(t, J = 7.3 Hz, 2H), 2.93 (dt, J = 8.0, 4.1 Hz, 1H), 2.49-2.23 (m, 3H),1.84 (q, J = 11.1, 8.9 Hz, 4H), 1.51-1.27 (m, 2H). 52 7 534 [M + H]⁺(300 MHz, MeOD-d₄) δ ppm: 8.31 (d, J = 2.6 Hz, 1H), 8.00-7.81 (m, 4H),7.75 (d, J = 1.7 Hz, 1H), 7.22-7.11 (m, 2H), 7.07-6.94 (m, 2H), 6.55(dd, J = 2.6, 1.8 Hz, 1H), 4.65 (t, J = 6.8 Hz, 1H), 4.35 (dq, J = 39.0,8.3 Hz, 2H), 4.13-3.91 (m, 2H), 3.22 (t, J = 7.3 Hz, 2H), 2.93 (dt, J =8.0, 4.1 Hz, 1H), 2.49-2.23 (m, 3H), 1.84 (q, J = 11.1, 8.9 Hz, 4H),1.51-1.27 (m, 2H). 53 7 535[M + H]⁺ (400 MHz, Methanol-d₄) δ 8.66 (d, J= 1.3 Hz, 1H), 8.12- 7.99 (m, 5H), 7.96 (d, J = 1.2 Hz, 1H), 7.07 (dd, J= 8.2, 5.1 Hz, 3H), 6.95 (t, J = 8.8 Hz, 3H), 5.18-5.05 (s, 1H),4.50-4.32 (m, 1H), 4.19-3.97 (m, 1H), 3.76-3.50 (m, 2H), 3.00-2.75 (m,3H), 2.46-2.30 (m, 2H), 2.00-1.60 (m, 5H), 1.30-0.95 (m, 8H), 1.08 (s,1H). 54 7 531 (M + 1) (400 MHz, Methanol-d₄) δ 8.64 (d, J = 1.2 Hz, 2H),8.11-7.96 (m, 8H), 7.93 (d, J = 1.2 Hz, 2H), 7.05 (ddd, J = 8.8, 5.3,2.0 Hz, 4H), 6.93 (ddd, J = 9.5, 7.7, 1.4 Hz, 4H), 4.99 (dd, J = 8.4,5.8 Hz, 1H), 4.38 (s, 1H), 4.09 (s, 1H), 3.99-3.91 (m, 1H), 3.63 (d, J =10.4 Hz, 1H), 3.52-3.43 (m, 1H), 3.36 (d, J = 2.6 Hz, 1H), 2.84- 2.68(m, 4H), 2.28 (dtd, J = 7.6, 4.7, 3.2 Hz, 2H), 2.06- 1.94 (m, 3H),1.95-1.59 (m, 26H), 1.10-0.91 (m, 4H) 55 7 546 [M + H]⁺ (300 MHz,MeOD-d₄) δ ppm: 8.68-8.53 (m, 1H), 8.14- 7.90 (m, 5H), 7.01 (ddt, J =8.3, 4.8, 2.4 Hz, 2H), 6.93- 6.74 (m, 2H), 5.00 (ddd, J = 21.2, 8.5, 5.5Hz, 1H), 4.22-3.66 (m, 2H), 3.52-3.32 (m, 1H), 3.22 (s, 2H), 3.02-2.80(m, 1H), 2.79-2.64 (m, 2H), 2.33-2.19 (m, 4H), 2.13-1.92 (m, 2H),1.91-1.67 (m, 4H), 1.67- 1.36 (m, 3H), 1.06-0.89 (m, 2H). 56 7 533 [M +H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.66 (d, J = 1.2 Hz, 1H), 8.11-7.92 (m,4H), 7.92 (s, 1H), 7.07-6.98 (m, 2H), 6.93-6.79 (m, 2H), 4.97 (ddd, J =25.1, 8.4, 5.5 Hz, 1H), 4.36 (dd, J = 8.1, 5.2 Hz, 2H), 4.19-3.69 (m,2H), 3.49-3.34 (m, 1H), 2.92 (td, J = 13.9, 2.5 Hz, 1H), 2.80- 2.61 (m,2H), 2.24 (ddd, J = 7.6, 4.6, 3.2 Hz, 1H), 2.06- 1.73 (m, 6H), 1.64 (m,3H), 1.12-0.77 (m, 2H). 57 7 574 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61(d, J = 1.3 Hz, 1H), 8.16-7.79 (m, 5H), 7.12-6.74 (m, 4H), 5.06 (dd, J =8.5, 5.3 Hz, 1H), 3.83-3.30 (m, 4H), 2.73 (tt, J = 8.2, 4.1 Hz, 2H),2.62 (t, J = 7.0 Hz, 2H), 2.54-2.38 (m, 2H), 2.32 (d, J = 4.6 Hz, 3H),2.30-2.18 (m, 1H), 1.94- 1.48 (m, 11H), 1.07-0.73 (m, 2H). 58 7 574 [M +H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61 (d, J = 1.3 Hz, 1H), 8.15-7.52 (m,5H), 7.12-6.69 (m, 4H), 4.81- 4.67 (m, 1H), 4.00-3.31 (m, 3H), 3.40-3.30(m, 1H), 3.25-3.18 (m, 1H), 2.75 (t, J = 7.2 Hz, 2H), 2.42 (d, J = 47.9Hz, 4H), 2.26 (t, J = 2.7 Hz, 4H), 1.96-1.49 (m, 11H), 1.11-0.84 (m,2H). 59 7 520[M + H]⁺ (400 MHz, MeOD-d₄) δ 8.07 (s, 1H), 8.10-7.90 (m,5H), 7.10-6.85 (m, 4H), 4.87 (s, 4H), 4.72-4.65 (m, 1H), 4.55- 4.45 (m,2H), 4.25-4.10 (m, 2H), 3.80-3.70 (m, 2H), 2.35- 60 7 588 [M + H]⁺ (300MHz, MeOD-d₄) δ ppm: 8.64 (s, 1H), 8.16-7.79 (m, 5H), 7.12-6.74 (m, 4H),5.06 (m, 1H), 3.83-3.45 (m, 4H), 3.19-3.02 (m, 2H), 2.89-2.60 (m, 6H),2.35-2.20 (m, 1H), 2.00-1.50 (m, 5H), 1.10-0.90 (m, 2H). 61 7 566[M +H]⁺ (400 MHz, Methanol-d₄) δ 8.54 (s, 1H), 8.05-7.80 (m, 5H), 7.05-6.80(m, 4H), 5.10-4.90 (m, 1H), 4.45-4.35 (m, 1H), 4.18-4.05 (m, 1H),3.55-3.30 (m, 1H), 3.00-2.85 (m, 1H), 2.90-2.60 (m, 2H), 2.25-1.50 (m,10H), 1.00-0.80 (m, 2H). 62 7 [M + H]⁺ (300 MHz, MeOD-d₄): δ ppm 9.07(s, 2H), 8.07-7.98 (m, 520 2H), 7.80-7.70 (m, 2H), 7.08-6.80 (m, 4H),5.15-4.95 (m, 1H), 4.50-4.26 (m, 1H), 4.10-3.95 (m, 1H), 3.90-3.80(m,1H), 3.40-3.20 (m, 3H), 2.78-2.74 (m, 2H), 2.32-2.28 (m, 1H), 1.94-1.77(m, 3H), 1.70-1.60 (m, 2H), 1.07-0.88 (m, 2H) 63 7 [M + H]⁺ (300 MHz,MeOD-d₄): δ ppm 9.07 (s, 2H), 8.07-7.98 (m, 534 2H), 7.85-7.70 (m, 2H),7.08-6.84 (m, 4H), 5.09-4.95 (m, 1H), 4.35-3.75 (m, 4H), 3.46-3.38 (m,2H), 3.22-3.15 (m, 2H), 2.95-2.90 (m, 4H), 2.44-2.40 (m, 1H), 1.90-1.83(m, 4H), 1.47-1.32 (m, 2H) 64 7 485 (300 MHz, MeOD-d₄): δ ppm 7.95-7.83(m, 2H), 7.25- 7.08 (m, 4H), 7.06-6.91 (m, 2H), 5.06-50.2(m, 1H),4.70-4.40(m, 2H), 3.60-3.55 (m, 4H), 3.28-3.11 (m, 4H), 3.18-3.05(m,2H), 2.92-2.88(m, 1H), 2.46-2.42 (m, 1H), 1.90-1.82 (m, 4H), 1.52-1.25(m, 5H) 65 7 [M + H]⁺ 463 (400 MHz, Methanol-d₄) δ 8.05-7.95 (m, 2H),7.89- 7.77(m, 2H), 7.11-7.00 (m, 2H), 7.00-6.88 (m, 2H), 4.70- 4.56 (m,1H), 4.23-4.10 (m, 1H), 4.00-3.90 (m, 1H), 3.78-3.70 (m, 1H), 3.69-3.60(m, 1H), 2.85-2.67 (m, 2H), 2.34-2.22 (m, 1H), 1.95-1.80 (m, 3H),1.78-7.50 (m, 2H), 1.30 (s, 6H), 1.11-1.00 (m, 1H), 1.00-0.89 (m, 1H).66 7 460 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 7.98 (m, 2H), 7.86 (m, 2H),7.19 (m, 2H), 7.03 (m, 2H), 4.74 (m, 1H), 4.65- 4.54 (m, 2H), 4.41-4.09(m, 2H), 3.75 (m, 1H), 3.25 (m, 2H), 2.95 (m, 1H), 2.45 (m, 1H), 1.95(m, 1H), 1.83 (m, 3H), 1.47-1.37 (m, 2H) 67 7 [M + H]⁺ 493 (400 MHz,Methanol-d₄) δ 8.05-7.93 (m, 2H), 7.89-7.77(m, 2H), 7.11-7.00 (m, 2H),7.00-6.88 (m, 2H), 4.68-4.50 (m, 1H), 4.50-4.30 (m, 1H), 4.30-4.16 (m,1H), 4.10-3.88 (m, 2H), 2.86-2.65 (m, 3H), 2.36-2.21 (m, 1H), 1.98-1.76(m, 3H), 1.76-1.53 (m, 2H), 1.25-1.11 (m, 6H), 1.11-1.00 (m, 1H),1.00-0.89 (m, 1H). 68 7 493 (M + 1) (CD₃OD, ppm): 8.10-7.97 (m, 2H),7.97-7.80 (m, 2H), 7.01-7.15 (m, 2H), 6.84-7.01 (m, 2H), 5.02-5.15 (m,1H), 3.99-4.26 (m, 1H), 3.72-3.98 (m, 1H), 3.45-3.72 (m, 1H), 3.10-3.30(m, 1H), 2.64-2.88 (m, 2H), 2.22-2.36 (m, 1H), 1.44-1.96 (m, 9H),1.19-1.38 (m, 3H), 0.92-1.11 (m, 2H). 69 7 507 (M + 1) (400 MHz,Methanol-d₄) δ 8.03-7.93 (m, 2H), 7.87- 7.78 (m, 2H), 7.09-6.99 (m, 2H),6.98-6.87 (m, 2H), 5.05 (dd, J = 8.7, 5.2 Hz, 1H), 4.19 (d, J = 13.2 Hz,1H), 3.89-3.73 (m, 1H), 3.54-3.36 (m, 1H), 3.22 (d, J = 2.8 Hz, 3H),2.99 (td, J = 12.6, 2.8 Hz, 1H), 2.82-2.67 (m, 2H), 2.26 (ddd, J = 7.5,4.4, 3.4 Hz, 1H), 1.93-1.73 (m, 5H), 1.72-1.35 (m, 4H), 1.18 (d, J = 9.1Hz, 3H), 1.08- 0.90 (m, 2H). 70 7 529 (M + 1) (400 MHz, Methanol-d₄) δ7.99 (dd, J = 8.4, 2.6 Hz, 2H), 7.87-7.79 (m, 2H), 7.69 (dd, J = 8.4,2.4 Hz, 1H), 7.49 (dd, J = 14.3, 1.9 Hz, 1H), 7.09-7.00 (m, 2H), 6.93(td, J = 8.7, 6.7 Hz, 2H), 6.29 (dt, J = 6.0, 2.1 Hz, 1H), 5.10 (dt, J =9.6, 4.9 Hz, 1H), 4.64 (t, J = 15.6 Hz, 1H), 4.53-4.43 (m,1H), 4.28(s,1H),3.44-3.30 (m,1H), 2.97-2.71 (m, 3H), 2.29 (dt, J = 7.1, 3.7 Hz,1H), 2.24-1.78 (m, 8H), 1.67 (t, J = 10.0 Hz, 2H), 1.09-0.92 (m, 2H). 717 546[M + H]⁺ (300 MHz, Methanol-d₄) δ 8.03-7.78 (m, 4H), 7.15-6.86 (m,4H), 5.10-5.00 (m, 1H), 3.85-3.60 (m, 3H), 3.60-3.50 (m, 1H), 3.20-2.10(m, 2H), 2.80-2.50 (m, 6H), 2.38-2.18 (m, 1H), 2.00-1.78 (m, 3H),1.75-1.52 (m, 2H), 1.10-0.85 (m, 2H). 72 7 [M + H]⁺ 530 (400 MHz,Methanol-d₄) δ 8.10-8.03 (m, 1H), 8.03-7.94 (m, 2H), 7.90-7.80 (m, 2H),7.80-7.70 (m, 1H), 7.10-7.00(m, 2H), 7.00-6.81 (m, 2H), 5.15-5.02 (m,1H), 4.80-4.51 (m, 1H), 4.40-4.20 (m, 1H), 350-3.48 (m, 1H), 3.18-2.88(m, 1H), 2.87-2.66 (m, 2H), 2.39-2.10 (m, 4H), 2.10-1.78 (m, 4H),1.78-1.67 (m, 2H), 1.10-0.87 (m, 2H). 73 7 571 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 8.67 (d, J = 7.2 Hz, 2H), 8.01-7.96 (m, 2H), 7.85 (d, J= 8.4 Hz, 2H), 7.05-7.03(m, 2H), 6.96-6.91 (m, 2H), 5.11-5.04(m, 1H),4.71-4.63(m, 1H), 4.61-4.50(m, 1H), 4.39-4.26(m, 1H), 3.31-3.30(m, 1H),2.91-2.75(m, 3H), 2.32-2.09(m, 4H), 1.96-1.83(m, 4H), 1.71-1.61(m, 2H),1.10-1.03(m, 1H), 0.98-0.96(m, 1H) 74 7 [M + H]⁺ 531; (300 MHz,Methanol-d₄) δ 8.02-7.90 (m, 2H), 7.90-7.86 (m, 2H), 7.26-7.18 (m, 2H),7.10-6.96 (m, 2H), 5.22-5.18 (m, 1H), 4.75-4.55 (m, 1H), 4.55-4.40 (m,1H), 4.30-4.10 (m, 1H), 3.25-3.23 (m, 1H), 3.12-2.90 (m, 3H), 2.50-2.41(m, 1H), 2.28-2.10 (m, 2H), 2.10-2.00 (m, 6H), 1.55-1.35 (m, 2H). 75 7[M + H]⁺ 492 (300 MHz, Methanol-d₄) δ 8.00-7.90 (m, 2H), 7.90-7.74 (m,2H), 7.11-7.00 (m, 2H), 7.00-6.82 (m, 2H), 5.10-4.95 (m, 1H), 3.85-4.42(m, 4H), 2.83-2.66 (m, 2H), 2.66-2.33 (m, 6H), 2.33-2.15 (m, 1H),1.94-1.71 (m, 3H), 1.71-1.48 (m, 2H), 1.20-1.05 (m, 3H), 1.05-0.80 (m,2H). 76 7 506.25 (300 MHz, Methanol-d₄) δ 7.94 (d, J = 8.2 Hz, 2H), 7.80(M + 1) (d, J = 8.2 Hz, 2H), 7.07-6.84 (m, 4H), 5.01 (dd, J = 8.6, 5.3Hz, 1H), 3.73-3.59 (m, 4H), 2.78-2.47 (m, 7H), 2.24 (dt, J = 7.6, 4.1Hz, 1H), 1.92-1.72 (m, 3H), 1.66-1.55 (m, 2H), 1.09-0.86 (m, 8H). 77 7508 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 7.97 (d, J = 8.3 Hz, 2H), 7.83(d, J = 8.3 Hz, 2H), 7.13-6.99 (m, 2H), 6.98-6.88 (m, 2H), 5.04 (m, 1H),3.81-3.60 (m, 5H), 3.60-3.49 (m, 1H), 2.75 (m, 2H), 2.55 (m, 6H), 2.27(m, 1H), 1.94- 1.72 (m, 3H), 1.63 (m, 2H), 1.08-0.91 (m, 2H). 78 7 [M +H]⁺ 522 (400 MHz, Methanol-d₄) δ 8.02-7.90 (m, 2H), 7.88-7.74 (m, 2H),7.14-7.00 (m, 2H), 7.00-6.86(m, 2H), 5.02- 5.10(m, 1H), 3.82-3.62 (m,3H), 3.62-3.51(m, 3H), 3.47- 3.43 (m, 3H), 2.81-2.68(m, 2H), 2.67-2.43(m, 6H), 2.35- 2.21 (m, 1H), 1.96-1.78 (m, 3H), 1.72-1.58 (m, 2H), 1.01-1.00(m, 1H), 1.00-0.90(m, 1H). 79 7 [M + H]⁺ 570 (300 MHz, Methanol-d₄)δ 8.02-7.90 (m, 2H), 7.90-7.80 (m, 2H), 7.13-7.00 (m, 2H), 7.00-6.86(m,2H), 5.00- 5.15(m, 1H), 3.86-3.45 (m, 5H), 3.45-3.36 (m, 1H), 3.07 (s,3H), 2.96-2.76 (m, 4H), 2.68-2.20 (m, 5H), 2.08-1.80 (m, 3H), 1.80-1.50(m, 2H), 1.25-0.92 (m, 2H). 80 7 [M + H]⁺ 506 (400 MHz, Methanol-d₄) δ8.20-7.90 (m, 2H), 7.90-7.68 (m, 2H), 7.20-7.02 (m, 2H), 7.02-6.80 (m,2H), 5.20-5.00 (m, 1H), 4.40-4.28 (m, 1H), 4.28-4.00 (m, 1H), 4.01-3.70(m, 2H), 3.65-3.42 (m, 4H), 3.00-2.85 (m, 2H), 2.58-2.25 (m, 1H),2.07-1.80 (m, 3H), 1.80-1.55 (m, 2H), 1.17 (t, J = 7.2 Hz, 3H),1.20-1.09(m, 1H), 1.09-1.00 (m, 1H). 81 7 501 [M + H]⁺ (300 MHz,MeOD-d₄) δ ppm: 7.96 (d, J = 8.5, Hz, 2H), 7.80 (d, J = 8.5 Hz, 2H),7.49-7.24 (m, 1H), 7.13-6.95 (m, 2H), 6.93-6.81 (m, 2H), 5.15-4.90 (m,1H), 4.79- 4.31 (m, 4H), 3.94-3.70 (m, 3H), 2.77 (t, J = 7.1 Hz, 2H),2.25 (m, 1H), 2.01-1.77 (m, 3H), 1.68 (m, 2H), 1.13-0.78 (m, 2H). 82 7565 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.05-7.95 (m, 3H), 7.87- 7.79 (m,2H), 7.04 (m, 2H), 6.97-6.86 (m, 2H), 5.11 (m, 1H), 4.87-4.64 (m, 2H),4.51-4.48 (m, 2H), 3.40 (s, 3H), 2.84-2.74 (m, 2H), 2.29 (m, 1H),2.00-1.83 (m, 3H), 1.70 (m, 2H), 1.10-0.91 (m, 2H). 83 7 477 [M + H]⁺(400 MHz, MeOD-d₄) δ ppm: 8.02-7.95 (m, 2H), 7.86- 7.75 (m, 2H),7.15-7.00 (m, 2H), 7.00-6.89 (m, 2H), 4.85-4.75(m, 4H), 4.75-4.65 (m,1H), 4.55-4.40 (m, 2H), 4.25-4.05 (m, 2H), 2.85-2.70 (m, 2H), 2.35-2.20(m, 1H), 21.95-1.52 (m, 5H), 1.10-0.95(m, 2H). 84 7 519[M + H]⁺ (400MHz, Methanol-d₄) δ 8.00-7.80(m, 4H), 7.15- 7.25(m, 2H), 7.0-7.1(m, 2H),5.1-5.2(m, 1H), 3.85- 3.95(m, 2H), 3.85-3.45(m, 5H), 3.33-3.21(m, 3H),300- 2.92(m, 1H), 2.4-2.5(m, 1H), 1.9-2.0(m, 1H), 1.75- 1.9(m, 5H),1.5-1.75(m, 4H), 1.35-1.55(m, 2H). 85 7 [M + H]⁺ (300 MHz, MeOD-d₄): δppm 8.85 (d, J = 4.9 Hz, 2H), 545 8.52-8.42 (m, 2H), 7.98-7.89 (m, 2H),7.37 (t, J = 4.9 Hz, 1H), 7.05-7.00 (m, 2H), 6.95-6.83 (m, 2H), 5.06-5.02(m, 1H), 3.78-3.62 (m, 4H), 3.56-3.50 (m, 1H), 2.76-2.72 (m, 2H),2.48-2.44(m, 6H), 2.28-2.22(m, 1H), 1.93-1.75 (m, 3H), 1.68-1.64(m, 2H),1.15-0.86 (m, 5H). 86 8 517 (M + 1) (CD₃OD, ppm): 8.87 (d, 2H), 8.49 (d,2H), 7.95 (d, 2H), 7.40 (t, 1H), 7.09-7.22 (m, 2H), 6.91-7.08 (m, 2H),5.00- 5.20 (m, 2H), 3.94-4.18 (m, 2H), 3.55-3.90 (m, 2H), 3.11-3.28 (m,6H), 2.84-2.98 (m, 1H), 2.32-2.52 (m, 1H) 1.73-2.00 (m, 4H), 1.29-1.41(m, 2H). 89 8 534 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.64 (s, 1H),8.10-7.91 (m, 5H), 7.05 (m, 2H), 6.92 (m, 2H), 5.14-5.03 (m, 1H), 4.42(m, 1H), 4.04 (m, 1H), 2.85-2.69 (m, 5H), 2.32-2.20 (m, 2H), 1.94-1.76(m, 3H), 1.71-1.62 (m, 2H), 1.17-0.91 (m, 8H). 90 8 532 [M + H]⁺ (300MHz, MeOD-d₄) δ ppm: 8.69-8.51 (m, 1H), 8.13- 7.93 (m, 4H), 7.95-7.85(m, 1H), 7.05-6.96 (m, 2H), 6.96-6.83 (m, 2H), 4.93 (t, J = 7.2 Hz, 1H),4.58-4.26 (m, 2H), 3.20-2.61 (m, 6H), 2.24 (dd, J = 7.7, 4.4 Hz, 1H),2.18-1.73 (m, 7H), 1.65 (t, J = 13.4 Hz, 2H), 1.06- 0.79 (m, 2H). 91 9549 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61 (s, 1H), 8.07-7.96 (m, 4H),7.91 (s, 1H), 7.16 (m, 2H), 6.99 (m, 2H), 5.11 (m, 1H), 4.45-4.18 (m,1H), 4.03-3.97 (m, 1H), 3.40- 2.78 (m, 5H), 2.63-2.42 (m, 2H), 1.94 1.25(m, 10H). 92 10 [M + H]⁺ 479 (400 MHz, Methanol-d₄) δ 8.02-7.90 (d, J =2.8 Hz, 2H), 7.90-7.80 (d, J = 2.8 Hz, 2H), 7.26-7.10 (m, 2H), 7.07-6.96(m, 2H), 5.18-5.06 (m, 1H), 4.30-3.80 (m, 3H), 3.58-3.36 (m, 1H),3.30-3.16 (m, 2H), 3.16-2.85 (m, 2H), 2.60-2.46 (m, 1H), 2.05-1.75 (m,6H), 1.62-1.30 (m, 4H). 93 10 532.3 (400 MHz, Methanol-d₄) δ 8.88 (d, J= 4.9 Hz, 2H), 8.50 (M + 1) (d, J = 8.3 Hz, 2H), 7.96 (dd, J = 8.5, 1.7Hz, 2H), 7.40 (t, J = 4.9 Hz, 1H), 7.04 (ddd, J = 8.6, 5.2, 2.6 Hz, 2H),6.97-6.85 (m, 2H), 5.11 (dt, J = 8.7, 4.5 Hz, 1H), 4.17 (d, J = 12.9 Hz,1H), 4.06-3.96 (m, 1H), 3.97-3.79 (m, 2H), 3.08 (ddd, J = 13.4, 10.2,3.2 Hz, 1H), 2.83- 2.72 (m, 2H), 2.28 (dd, J = 6.8, 3.8 Hz, 1H),1.92-1.78 (m, 6H), 1.74-1.52 (m, 3H), 1.46 (td, J = 10.1, 9.4, 5.0 Hz,1H), 1.09-0.91 (m, 2H). 94 10 [M + H]⁺ 478 (300 MHz, MeOD-d₄):δ7.99-7.89 (m, 2H), 7.84-7.75 (m, 2H), 7.07-6.84 (m, 4H), 5.03-4.95 (m,1H), 3.81-3.47 (m, 4H), 2.75-2.70 (m, 2H), 2.53-2.18 (m, 8H), 1.92-1.67(m, 3H), 1.62-1.57 (m, 2H), 1.07-0.86 (m, 2H). 95 10 531 [M + H]⁺ (300MHz, MeOD-d₄) δ ppm: 8.04 (s, 4H), 7.33-7.07 (m, 2H), 7.07-6.85 (m, 2H),5.07 (t, J = 6.6 Hz, 1H), 4.40-3.60 (m, 3H), 3.22 (d, J = 13.0 Hz, 7H),3.14 (s, 3H), 2.90 (s, 4H), 2.52-2.24 (m, 1H), 1.87 (q, J = 11.0 Hz,4H), 1.59-1.34 (m, 1H), 1.30 (dd, J = 15.7, 8.6 Hz, 1H). 96 10 560(M + 1) (CD₃OD, ppm): 7.97-8.12 (m, 2H), 7.74-7.97 (m, 2H), 6.98-7.13(m, 2H), 6.84-6.98 (m, 2H), 4.96-5.12 (m, 1H), 3.42-3.85 (m, 4H),2.60-2.80 (m, 8H), 2.34-2.60 (m, 4H), 2.12-2.34 (m, 4H), 1.72-1.93 (m,3H), 1.50-1.72 (m, 2H), 0.97-1.05 (m, 2H) 97 10 518 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 7.97-7.89 (m, 2H), 7.62- 7.53 (m, 2H), 7.29 (t, J = 2.2Hz, 2H), 7.10-7.00 (m, 2H), 6.98-6.87 (m, 2H), 6.32 (t, J = 2.2 Hz, 2H),5.06 (m, 1H), 3.78 (m, 2H), 3.66 (m, 1H), 3.58 (m, 1H), 2.84- 2.69 (m,2H), 2.55-2.37 (m, 4H), 2.34-2.23 (m, 4H), 1.94-1.73 (m, 3H), 1.66 (m,2H), 1.09-0.91 (m, 2H). 98 10 520 (M + 1) (CD₃OD, ppm): 8.26-8.11 (m,2H), 8.10-7.91 (m, 4H), 7.13-7.00 (m, 2H), 7.00-6.81 (m, 2H), 5.13-4.98(m, 1H), 3.81-3.68 (m, 2H), 3.61-3.51 (m, 1H), 2.85-2.68 (m, 2H),2.60-2.38 (m, 4H), 2.38-2.18 (m, 4H), 1.99-1.77 (m, 3H), 1.77-1.60 (m,2H), 1.36-1.28 (m, 1H), 1.08-0.93 (m, 2H) 99 10 534 (M + 1) (CD₃OD,ppm): 8.07 (s, 1H), 7.91 (s, 4H), 7.09-6.99 (m, 2H), 6.99-6.88 (m, 2H),5.12-5.02 (m, 1H), 4.28 (s, 3H), 3.85-3.65 (m, 3H), 3.58-3.48 (m, 1H),2.82-2.72 (m, 2H), 2.58-2.40 (m, 4H), 2.37-2.28 (s, 3H), 2.37-2.27 (m,1H), 1.97-1.77 (m, 3H), 1.77-1.59 (m, 2H), 1.08-0.91 (m, 2H). 100 10 520[M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.20(s, 1H), 8.20 (s, 1H),8.04-8.01(m, 2H), 7.96-7.94(m, 2H), 7.05-7.02(m, 2H), 6.94-6.89(m, 2H),5.08-5.04(m, 1H), 3.81-3.62(m, 3H), 3.59-3.51(m, 1H), 2.78-2.73(m, 2H),2.55-2.36(m, 4H), 2.29-2.25(m, 4H), 1.89-1.86(m, 3H), 1.66-1.65(m, 2H),1.04-1.02(m, 1H), 0.96-0.94(m, 1H) 101 10 534 (M + 1) (CD₃OD, ppm): 8.48(s, 1H), 8.13 (m, 2H), 7.94 (m, 2H), 7.15-7.01 (m, 2H), 6.99-6.86 (m,2H), 5.13-5.02 (m, 1H), 4.02 (s, 3H), 3.90-3.48 (m, 5H), 2.85-2.68 (m,2H), 2.60-2.38 (m, 4H), 2.38-2.28 (m, 4H), 2.00-1.60 (m, 5H), 1.12-0.88(m, 2H). 102 10 [M + H]⁺ 521 (300 MHz, Methanol-d₄) δ 8.20-8.10 (m, 2H),8.10-7.95 (m, 2H), 7.28-7.10(m, 2H), 7.10-6.86(m, 2H), 5.14- 5.06(m,1H), 4.48-3.60(m, 3H), 3.56-3.31(m, 3H), 3.26- 3.06(m, 3H), 2.99-2.88(m,4H), 2.53-2.40(m, 1H), 2.03- 1.78(m, 4H), 1.53-1.40(m, 1H), 1.40-1.25(m,2H). 103 10 531 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.85(s, 2H), 8.48(d,J = 8.4 Hz, 2H), 8.05-7.90 (m, 2H), 7.55-7.40 (m, 1H), 7.10- 6.98 (m,2H), 6.98-6.80 (m, 2H), 5.15-5.00 (m, 1H), 3.88-3.45 (m, 4H), 2.88-2.70(m, 2H), 2.60-2.35 (m, 4H), 2.35-2.20 (m, 4H), 1.96-1.75 (m, 3H),1.75-1.62 (m, 2H), 1.10-0.88 (m, 2H) 104 10 531 (M + 1) (CD₃OD, ppm):9.18 (s, 1H), 8.47 (dd, 2H), 7.45-7.61 (m, 3H), 6.69-7.11 (m, 2H),6.78-6.99 (m, 2H), 5.01-5.12 (m, 1H), 3.58-3.88 (m, 3H), 3.47-3.56 (m,1H), 2.72-2.93 (m, 2H), 2.34-2.58 (m, 5H), 2.19-2.33 (m, 3H), 1.92-2.05(m, 1H), 1.74-1.92 (m, 2H), 1.55-1.74 (m, 2H), 0.97-1.15 (m, 2H). 105 10533 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.36-8.31 (m, 1H), 7.89- 7.79 (m,3H), 7.30 (m, 1H), 7.20 (m, 1H), 7.07-6.97 (m, 2H), 6.95-6.84 (m, 2H),5.10 (m, 1H), 3.94 (s, 3H), 3.86-3.60 (m, 3H), 3.54 (m, 1H), 2.85-2.69(m, 2H), 2.56-2.35 (m, 4H), 2.33-2.22 (m, 4H), 1.97-1.76 (m, 3H), 1.67(m, 2H), 1.08-0.89 (m, 2H). 106 10 559 (M + 1) (CD₃OD, ppm): 8.85 (d,2H), 8.48 (d, J = 8.4 Hz, 2H), 7.95 (d, J = 8.7 Hz, 2H), 7.45 (dd, J =4.8 Hz, 5.1 Hz, 1H), 7.14-6.96 (m, 2H), 6.96-6380 (m, 2H), 5.20-5.00 (m,1H), 3.90-3.45 (m, 4H), 2.85-2.66 (m, 3H), 2.66-2.40 (m, 4H), 2.38-2.12(m, 1H), 2.00-1.75 (m, 3H), 1.75-1.46 (m, 2H), 1.15-0.80 (m, 7H). 107 10574 (M + 1) (CD₃OD, ppm): 8.64 (s, 1H), 7.91-8.12 (m, 5H), 7.01- 7.11(m, 2H), 6.89-7.00 (m, 2H), 5.03-5.12 (m, 1H), 3.44-3.99 (m, 9H),2.71-2.85 (m, 2H), 2.25-2.38 (m, 1H), 1.82-1.98 (m, 3H), 1.62-1.76 (m,2H), 1.27-1.38 (m, 1H), 0.92-1.11 (m, 2H). 108 10 504 [M + H]⁺ (300 MHz,MeOD-d₄) δ ppm: 8.04-7.91 (m, 2H), 7.80 (d, J = 8.3 Hz, 2H), 7.08-6.96(m, 2H), 6.95-6.85 (m, 2H), 5.00 (dd, J = 8.6, 5.5 Hz, 1H), 4.53-4.35(m, 1H), 3.98 (ddt, J = 64.0, 15.0, 5.1 Hz, 1H), 3.78-3.41 (m, 1H),3.11-2.90 (m, 6H), 2.81-2.64 (m, 2H), 2.24 (dt, J = 6.8, 3.5 Hz, 1H),2.08-1.52 (m, 9H), 0.96 (ddt, J = 23.4, 7.0, 5.4 Hz, 2H). 109 11 498[M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.11-8.09 (m, 2H), 7.92- 7.89 (m,2H), 7.18-7.16 (m, 2H), 7.03-6.99 (m, 2H), 5.14 (m, 1H), 4.22-3.88 (m,3H), 3.45 (m, 1H), 3.27- 3.18 (m, 5H), 2.93 (m, 1H), 2.45 (m, 1H),1.83-1.82 (m, 6H), 1.45 (m, 4H) 110 12 520 [M + H]⁺ (400 MHz, MeOD-d₄) δppm: δ 8.64 (m, 1H), 8.10- 7.96 (m, 4H), 7.93 (m, 1H), 7.09-7.01 (m,2H), 6.96- 6.86 (m, 2H), 5.07 (m, 1H), 3.77 (m, 2H), 3.67 (m, 1H), 3.54(m, 1H), 2.81 (m, 2H), 2.55-2.38 (m, 4H), 2.32 (m, 4H), 1.95-1.68 (m,4H), 1.36-1.26 (m, 1H), 1.12- 0.94 (m, 2H). 111 13 521 (M + 1) (400 MHz,Methanol-d₄) δ 8.63 (d, J = 1.3 Hz, 1H), 8.07-8.05 (d, J = 7.2 Hz, 2H),8.00-7.98 (d, J = 7.2 Hz, 2H), 7.93 (d, J = 1.2 Hz, 1H), 7.09-7.00 (m,2H), 6.98- 6.87 (m, 2H), 5.11 (m, 1H), 4.16-3.80 (m, 3H), 3.39- 3.31 (m,1H), 2.77 (m, 2H), 2.27 (m, 1H), 2.05-1.81 (m, 6H), 1.75-1.43 (m, 4H),1.09-0.91 (m, 2H). 112 13 519 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: δ 8.26(s, 1H), 8.01 (d, J = 2 Hz, 2H), 7.68(d, J = 2 Hz, 2H), 7.67(s, 1H),7.18 (m, 1H), 7.10-7.00 (m, 2H), 6.99-6.87 (m, 2H), 5.06 (m, 1H),3.85-3.62 (m, 3H), 3.54 (m, 1H), 2.85-2.69 (m, 2H), 2.56-2.38 (m, 4H),2.34-2.23 (m, 4H), 1.89-1.70 (m, 3H), 1.60-1.56 (m, 2H), 1.09-0.91 (m,2H). 113 13 499.4 (CD₃OD, ppm): 7.95-7.88 (m, 2H), 7.25-7.10 (m, 2H),(M + 1) 7.10-6.98 (m, 2H), 6.98-6.82 (m, 2H), 5.10-4.95 (m, 1H),3.87-3.40 (m, 4H), 2.80-2.63 (m, 3H), 2.63-2.46 (m, 4H), 2.28-2.15 (m,1H), 1.91-1.70 (m, 3H), 1.70-1.52 (m, 2H), 1.10-0.90 (m, 7H). 114 13 547[M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.20 (d, J = 1.6 Hz, 1H), 8.37 (dd, J= 2.4 Hz, 6 Hz, 1H), 8.00-7.90(m, 1H), 7.20- 7.00(m, 2H), 7.00-6.89 (m,2H), 5.15-5.05 (m, 1H), 3.80- 3.50 (m, 4H), 3.20-3.05 (m, 2H), 2.85-2.60(m, 6H), 2.30-2.20 (m, 1H), 2.00-1.78 (m, 3H), 1.78-1.60 (m, 2H),1.10-0.95 (m, 2H). 115 13 557 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:7.77-7.68 (m, 2H), 7.22- 7.12 (m, 2H), 7.06-6.95 (m, 2H), 6.62-6.53 (m,2H), 5.10 (m, 1H), 4.43 (m, 1H), 4.22 (m, 1H), 3.88 (m, 1H), 3.73-3.63(m, 1H), 3.38-3.10 (m, 10H), 2.92 (m, 1H), 2.44 (m, 1H), 2.11-1.99 (m,4H), 2.00-1.75 (m, 4H), 1.45 (m, 1H), 1.39-1.29 (m, 1H). 116 13 501[M +H]⁺ (400 MHz, Methanol-d₄) δ 8.26 (s, 1H), 8.05-7.98 (m, 2H), 7.75-7.65(m, 3H), 7.19 (s, 1H), 7.10-7.00 (m, 2H), 7.00-6.90 (m, 2H), 4.90-4.10(m, 5H), 3.88-3.76 (m, 1H), 2.85-2.70 (m, 2H), 2.36-2.21 (m, 1H),2.00-1.55 (m, 5H), 1.15-0.95 (m, 2H). 117 14 520[M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: δ 8.64 (s, 1H), 8.11- 7.96 (m, 4H), 7.93 (m, 1H), 7.05(m, 2H), 6.98-6.88 (m, 2H), 4.68-3.75 (m, 5H), 3.27-3.16 (m, 1H), 2.78(m, 2H), 2.28 (m, 1H), 2.21 (d, J = 5.9 Hz, 6H), 1.95- 1.79 (m, 3H),1.66 (m, 2H), 1.09-0.91 (m, 2H). 118 14 584 (M + 1) (CD₃OD, ppm):8.69-8.91 (m, 2H), 8.38-8.45 (m, 2H), 7.89-8.00 (m, 2H), 7.33-7.44 (m,1H), 6.98-7.07 (m, 2H), 6.79-6.96 (m, 2H), 4.96-5.12 (m, 1H), 3.73-3.95(m, 3H), 3.44-3.58 (m, 4H), 2.68-2.72 (m, 2H), 2.18-2.31 (m, 1H),1.77-1.93 (m, 3H), 1.57-1.71 (m, 2H), 0.80-1.08 (m, 3H) 119 14 522[M +H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.85(s, 1H), 8.10-8.08 (m, 2H), 8.01-7.99(m, 2H), 7.07-7.03 (m, 2H), 6.94- 6.90 (m, 2H), 5.12 (m, 1H), 4.09 (m,3H), 3.55-3.04 (m, 3H), 2.91 (m, 2H), 2.32 (m, 1H), 2.03-1.40 (m, 9H),1.02 (m, 2H). 120 14 [M + H]⁺ 521 (300 MHz, MeOD-d₄): δ9.82 (s, 1H),8.11-7.92 (m, 4H), 7.08-6.83 (m, 4H), 5.10-5.03 (m, 1H), 3.79-3.57 (m,4H), 2.78-2.72 (m, 2H), 2.53-2.19 (m, 8H), 1.93- 1.74 (m, 3H),1.68-1.62(m, 2H), 1.08-0.87 (m, 2H). 121 14 478 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 8.05-7.90 (m, 2H), 7.90- 7.70 (m, 2H), 7.10-7.00 (m,2H), 7.00-6.86 (m, 2H), 5.05-4.90 (m, 1H), 4.50-4.25(m, 1H), 4.10-3.90(m, 1H), 3.90-3.62 (m, 1H), 3.45-3.37 (m, 1H), 3.37-3.30 (m, 1H),3.28-3.18 (m, 1H), 2.80-2.60 (m, 2H), 2.30-2.18 (m, 1H), 1.95-1.50 (m,5H), 1.10-0.85 (m, 2H) 122 14 478 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:9.20-8.85(m, 1H), 8.20- 8.00 (m, 2H), 8.00-7.96 (m, 2H), 7.15-6.88 (m,4H), 5.05-4.65 (m, 1H), 4.26-3.80 (m, 3H), 3.75-3.60 (m, 1H), 3.45-3.38(m, 1H), 3.30-3.20 (m, 1H), 2.95-2.80 (m, 3H), 2.70-2.58 (m, 2H),2.20-2.12 (m, 1H), 1.90-1.67 (m, 3H), 1.60-1.36 (m, 2H), 1.00-0.80 (m,2H) 123 14 546 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.64 (s 1H), 8.11-7.91(m, 5H), 7.04 (m, H), 6.98-6.87 (m, 2H), 5.11-5.00 (m, 1H), 4.54 (m,1H), 4.01-3.51(m, 1H), 3.02 (m, 6H), 2.78 (m, 2H), 2.28 (m, 1H),2.04-1.60 (m, 9H), 1.00 (m, 2H). 124 14 557 [M + H]⁺ (300 MHz, MeOD-d₄)δ ppm: 8.86 (dd, J = 4.9, 1.1 Hz, 2H), 8.47 (dd, J = 8.4, 1.7 Hz, 2H),7.94 (d, J = 8.4 Hz, 2H), 7.37 (t, J = 4.9 Hz, 1H), 7.02 (dd, J = 8.6,5.4 Hz, 2H), 6.89 (t, J = 8.5 Hz, 2H), 5.05 (dd, J = 8.3, 5.2 Hz, 1H),4.47 (d, J = 27.3 Hz, 1H), 4.16-3.82 (m, 1H), 3.81- 3.47 (m, 1H), 3.02(dq, J = 13.2, 6.4 Hz, 6H), 2.84- 2.70 (m, 2H), 2.26 (dd, J = 7.1, 3.9Hz, 1H), 2.10-1.56 (m, 9H), 1.10-0.84 (m, 2H) 125 14 463 [M + H]⁺ (400MHz, MeOD-d₄) δ 7.98 (d, J = 4.8 Hz, 2H), 7.83 (d, J = 2 Hz, 2H),7.10-7.00 (m, 2H), 7.00-6.90 (m, 2H), 4.96-4.90 (m, 1H), 4.77-4.62 (m,1H), 4.80-4.65 (m, 1H), 4.60-4.45 (m, 2H), 4.45-4.20 (m, 2H), 2.85-2.72(m, 2H), 2.35-2.22 (m, 1H), 2.00-1.75 (m, 3H), 1.70-1.58 (m, 2H),1.10-0.90 (m, 2H) 126 15 522 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:8.03-7.95 (d, J = 4.8 Hz, 2H), 7.89-7.81 (d, J = 4.8 Hz, 2H), 7.23-7.13(m, 2H), 7.07-6.96 (m, 2H), 5.08 (m, 1H), 4.09 (m, 4H), 3.91 (m, 1H),3.74 (m, 1H), 3.40 (m, 3H), 3.25 (m, 3H), 2.93 (m, 1H), 2.47 (m, 1H),2.00-1.77 (m, 4H), 1.48 (m, 1H), 1.35 (m, 1H). 127 16 [M + H]⁺ (400 MHz,Methanol-d₄) δ 8.64 (s, 1H), 8.10-7.99(m, 654; 4H), 7.99-7.92 (m, 1H),7.39-7.22 (m, 5H), 7.15-6.90 (m, 2H), 6.90-6.75 (m, 2H), 5.21-5.13 (m,3H), 3.78-3.52 (m, 6H), 3.49-3.40 (m, 1H), 2.75-2.64 (m, 1H), 2.55-2.40(m, 4H), 2.40-2.25 (m, 3H), 2.25-2.10 (m, 1H), 1.93-65 (m, 4H),1.52-1.10 (m, 3H). 128 17 [M + H]⁺593 (300 MHz, MeOD-d₄) δ ppm:7.89~7.87(m, 2H), 7.70~7.62(m, 4H), 7.46~7.35(m, 3H), 7.04~6.99(m, 2H),6.89~6.86(, .2H), 5.07~5.02(m, 1H), 3.92~3.86(m, 2H), 3.72~3.65(m, 1H),3.51~3.45(m, 1H), 3.38~3.32(m, 2H), 3.28~3.25(m, 1H), 3.18~3.07(m, 1H),2.83(s, 3H), 2.77~2.72(m, 2H), 2.26~2.24(m, 1H), 1.89~1.82(m, 3H),1.66~1.64(m, 2H), 1.03~0.92(m, 2H). 129 18 532 [M + H]⁺ (400 MHz,Methanol-d₄) δ 8.63 (s, 1H), 8.06-8.03 (m, 2H), 8.03-7.98 (m, 2H),7.97-7.93(m, 1H), 6.96-6.94 (m, 2H), 6.76-6.74 (m, 2H), 5.08-5.07 (m,1H), 3.76-3.63 (m, 6H), 3.57-3.51 (m, 1H), 2.80-2.74 (m, 2H), 2.56-2.38(m, 4H), 2.31 (s, 3H), 2.26-2.20 (m, 1H), 1.89-1.81 (m, 3H), 1.72-1.64(m, 2H), 0.99-0.89 (m, 2H) 130 19 [M + H]⁺ 538 (400 MHz, Methanol-d₄) δ8.67 (s, 1H), 7.99-7.82 (m, 4H), 7.05 (ddd, J = 8.4, 5.3, 2.6 Hz, 2H),6.98-6.86 (m, 2H), 5.10 (m, 1H), 3.83-3.50 (m, 4H), 2.84-2.68 (m, 2H),2.56-2.36 (m, 4H), 2.59 (s, 3H) 2.25-2.23 (m, 1H), 1.97-1.79 (m, 2H),1.79-1.70 (m, 1H), 1.69- 1.60 (m, 2H), 1.08-0.91 (m, 2H). 131 20 548[M +H]⁺ (400 MHz, Methanol-d₄) δ 8.66 (s, 1H), 8.12-7.99 (m, 4H), 7.96 (s,1H), 7.15-7.02(m, 2H), 7.02-6.88 (m, 2H), 5.17-5.01 (s, 1H), 4.20-3.63(m, 3H), 3.48-3.36 (m, 1H), 2.78 (s, 2H), 2.51-2.10 (s, 5H), 1.98-1.79(m, 3H), 1.79-1.51 (m, 2H), 1.12-0.89 (m, 2H) 132 21 520 [M + H]⁺ (400MHz, MeOD-d₄) δ ppm: 8.65(s, 1H), 8.06-8.01 (m, 2H), 7.98-7.93(m, 2H),7.93(s, 1H), 7.05-7.01(m, 2H), 6.92-6.87(m, 2H), 5.08-5.05(m, 1H),3.81-3.62(m, 3H), 3.57-3.50(m, 1H), 2.78-2.75(m, 2H), 2.55-2.49(m, 4H),2.35-2.26(m, 4H), 1.89-1.86(m, 3H), 1.84-1.82(m, 2H), 1.04-1.02(m, 1H),0.96-0.95(m, 1H) 133 21 520 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.65(s,1H), 8.15-7.94 (m, 5H), 7.11-7.05(m, 2H), 6.96-6.87(m, 2H), 5.15-5.05(m,1H), 3.87-3.47(m, 4H), 2.87-2.72(m, 2H), 2.55-2.40(m, 4H), 2.35-2.32(m,3H), 2.32-2.27 (m, 1H), 1.99-1.76(m, 3H), 1.76-1.52 (m, 2H),1.14-1.02(m, 1H), 1.02-0.91(m, 1H) 134 22 531 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 9.19 (d, J = 1.6 Hz, 1H), 8.73 (dd, J = 2.5, 1.5 Hz,1H), 8.60 (d, J = 2.5 Hz, 1H), 8.26-8.18 (m, 2H), 8.05-7.98 (m, 2H),7.23-7.13 (m, 2H), 7.07-6.94 (m, 2H), 5.12 (m, 1H), 4.68-4.48 (m, 2H),3.57 (s, 3H), 3.36-3.20 (m, 2H), 3.13 (m, 3H), 2.98-2.89 (m, 4H), 2.47(m, 1H), 2.03-1.80 (m, 4H), 1.49 (m, 1H), 1.35 (m, 1H). 135 23 531(M + 1) (CD₃OD, ppm): 9.20 (s, 1H), 9.13 (s, 2H), 8.02 (d, J = 6.3 Hz,2H), 7.84 (d, J = 6.9 Hz, 2H), 7.12-7.00 (m, 2H), 700-6.86 (m, 2H),5.15-5.05 (m, 1H), 3.86-3.52 (m, 4H), 2.85-2.73 (m, 2H), 2.55-2.45 (m,3H), 2.30 (s, 3H), 2.30- 2.22 (m, 1H), 1.92-1.85 (m, 2H), 1.80-1.60 (m,2H), 1.40-0.90 (m, 4H). 136 23 531 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:9.21-9.20(m, 1H), 8.28- 8.21(m, 3H), 8.05-8.03(m, 2H), 7.87-7.83(m, 1H),7.08- 7.05(m, 2H), 6.96-6.92(m, 2H), 5.10-5.09(m, 1H), 3.85- 3.52(m,4H), 2.80-2.78(m, 2H), 2.54-2.43(m, 4H), 2.33 (s, 3H), 2.30-2.29(m, 1H),1.94-1.82(m, 3H), 1.73- 1.65(m, 2H), 1.07-1.05(m, 1H), 0.98-0.97(m, 1H),137 24 523.3 (400 MHz, Methanol-d₄) δ 8.64 (d, J = 1.3 Hz, 1H), 8.11-(M + 1) 7.91 (m, 5H), 7.04 (ddd, J = 8.5, 5.3, 2.6 Hz, 2H), 6.97- 6.86(m, 2H), 5.07 (dd, J = 8.7, 5.2 Hz, 1H), 3.87-3.40 (m, 4H), 2.77 (ddd, J= 11.4, 8.6, 5.1 Hz, 2H), 2.59- 2.35 (m, 4H), 2.27 (ddd, J = 7.5, 4.4,3.3 Hz, 1H), 1.85 (dddd, J = 28.2, 14.0, 7.4, 4.4 Hz, 3H), 1.74-1.59 (m,2H), 1.08-0.90 (m, 2H). 138 25 528 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm:8.60 (s, 1H), 8.17-7.84 (m, 5H), 7.11-7.01 (m, 2H), 6.95-6.77 (m, 2H),5.03 (m, 1H), 2.75 (m, 2H), 2.38-2.10 (m, 4H), 1.95-1.67 (m, 3H),1.70-1.58 (m, 2H), 1.14-0.80 (m, 2H). 139 565 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: δ8.33(s, 1H), 7.99-7.96(d, J = 8.8 Hz, 2H), 7.87-7.85(d,2H), 7.76(s, 1H), 7.05- 7.02(m, 2H), 6.92-6.87(m, 2H), 6.57-6.56(m, 1H),5.14- 5.11(m, 1H), 4.19-4.16(m, 1H), 3.57-3.41(m, 1H), 3.12- 2.95(m,1H), 2.80-2.72(m, 2H), 2.29-1.78(m, 8H), 1.71- 1.61(m, 2H), 1.06-1.01(m,1H), 0.96-0.92(m, 1H). 140 519 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: δ 8.25(s, 1H), 8.01 (d, J = 2 Hz, 2H), 7.68(d, J = 2 Hz, 2H), 7.67(s, 1H),7.18 (m, 1H), 7.10-7.00 (m, 2H), 6.99-6.87 (m, 2H), 5.08 (m, 1H),3.85-3.62 (m, 3H), 3.54 (m, 1H), 2.85-2.69 (m, 2H), 2.56-2.38 (m, 4H),2.34-2.23 (m, 4H), 1.89-1.70 (m, 3H), 1.60-1.56 (m, 2H), 1.09-0.91 (m,2H). 141 2 607[M + H]⁺ (400 MHz, Methanol-d₄) δ 7.96 (dd, J = 8.5, 1.8Hz, 2H), 7.76 (d, J = 8.1 Hz, 2H), 7.73-7.66 (m, 2H), 7.49 (t, J = 7.6Hz, 2H), 7.41 (t, J = 7.3 Hz, 1H), 7.26-7.19 (m, 2H), 7.10-7.02 (m, 2H),5.11 (dd, J = 8.6, 5.5 Hz, 1H), 4.01 (dd, J = 26.1, 13.6 Hz, 2H), 3.66(t, J = 11.0 Hz, 1H), 3.56-3.36 (m, 3H), 3.22 (t, J = 7.8 Hz, 3H),3.18-3.08 (m, 1H), 2.97 (dt, J = 7.8, 4.0 Hz, 1H), 2.88 (s, 3H), 2.47(ddd, J = 10.4, 6.7, 3.6 Hz, 1H), 1.86 (dtt, J = 39.1, 16.6, 7.5 Hz,4H), 1.68-1.43 (m, 3H), 1.39 (q, J = 7.1 Hz, 1H). 142 26 534 [M + H]⁺(400 MHz, CD₃OD-d₄) δ ppm: 8.67 (d, J = 1.2 Hz, 1H), 8.18-8.01 (m, 4H),7.96 (d, J = 1.3 Hz, 1H), 7.29-7.15 (m, 2H), 7.13-6.97 (m, 2H), 5.06 (m,1H), 4.8- 4.3(m, 2H), 3.9-2.9 (m, 12H), 2.50 (m, 1H), 2.03-1.71 (m, 4H),1.69-1.33 (m, 4H). 143 27 492 [M + H]⁺ (300 MHz, CD₃OD-d₄) δ ppm:8.02-7.88 (d, J = 11.6 Hz, 2H), 7.88-7.70 (d, J = 11.6 Hz, 2H), 7.07-6.81 (m, 4H), 4.99 (m, 1H), 3.73-3.69 (m, 2H), 3.64- 3.38 (m, 2H), 2.68(t, J = 7.1 Hz, 2H), 2.57-2.34 (m, 4H), 2.30-2.16 (m, 4H), 1.97-1.67 (m,3H), 1.49-1.3 (m, 4H), 1.08-0.82 (m, 2H). 144 28 545 [M + H]⁺ (300 MHz,CD₃OD-d₄) δ ppm: 8.85 (d, J = 4.9 Hz, 2H), 8.48 (d, J = 8.3 Hz, 2H),7.94 (d, J = 8.3 Hz, 2H), 7.38 (t, J = 4.9 Hz, 1H), 7.03 (t, J = 8.7 Hz,2H), 6.91 (t, J = 8.7 Hz, 2H), 5.03 (m, 1H), 4.60-4.55 (m, 2H),3.82-3.44 (m, 4H), 2.72 (t, J = 7.1 Hz, 2H), 2.49-2.30(m, 4H), 2.29 (m,4H), 1.98-1.73 (m, 3H), 1.67-1.35 (m, 4H), 1.12- 0.86 (m, 2H). 145 29[M + H]+ (400 MHz, CDCl₃): δ7.87~7.82 (m, 2H), 7.17~7.07 536 (m, 4H),6.99~6.95 (m, 2H), 5.70~5.65 (m, 1H), 3.81~3.64 (m, 10H), 3.42~3.50 (m,1H), 3.39~3.33 (m, 3H), 2.22~2.30 (m, 1H), 1.10~1.20 (m, 1H), 1.00 (s,3H), 0.90~0.87 (m, 1H) 146 30 568 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm:8.00-7.91(m, 2H), 7.62- 7.48(m, 2H), 7.12-7.00(m, 2H), 7.00-6.81 (m,2H), 6.09 (s, 1H), 5.18-5.08 (m, 1H), 4.50-4.28 (m, 2H), 4.00-3.81 (m,1H), 3.78-3.59 (m, 1H), 3.58-3.36 (m, 1H), 3.21-3.03 (m, 3H), 2.91-2.78(m, 2H), 2.34-2.25 (m, 4H), 2.10-1.94 (m, 2H), 1.94-1.80 (m, 1H),1.10-0.90(m, 2H) 147 2 515 [M + H]⁺ (300 MHz, Methanol-d₄) δ 8.07-7.91(m, 6H), 7.58 (m, 3H), 7.06-6.83 (m, 4H), 5.08 (dd, J = 8.5, 5.3 Hz,1H), 4.36 (m, 2H), 3.89 (m, 2H), 3.66 (s, 1H), 3.39 (s, 1H), 3.10 (s,3H), 2.81 (t, J = 6.7 Hz, 2H), 2.25 (dd, J = 7.2, 3.9 Hz, 1H), 2.04-1.87(m, 2H), 1.06-0.87 (m, 2H) 148 31 [M + H]+567 (300 MHz, CD₃OD): δ(ppm):7.98-7.96 (d, 1H, J = 6 HZ), 7.34~7.32 (d, 2H, J = 6 Hz), 7.19-7.15 (t,2H, J = 6 Hz), 7.04-7.00 (t, 2H, J = 6 Hz), 5.81 (s, 2H), 5.24-5.19 (m,1H), 4.35-4.27 (m, 1H), 4.16-4.05 (m, 1H), 3.87-3.85 (m, 2H), 3.33-3.28(m, 2H), 3.16-3.10 (m, 4H), 2.99-2.95 (m, 1H), 2.48-2.43 (m, 1H),2.29-2.10 (m, 2H), 1.98 (s, 6H), 1.50-1.40 (m, 1H), 1.38-1.33 (m, 1H)150 32 550 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.05-7.94 (m, 2H), 7.85-7.75 (m, 2H), 7.74-7.64 (m, 2H), 7.55-7.46 (m, 2H), 7.46-7.39 (m, 1H),7.30-7.18 (m, 2H), 7.11-7.01 (m, 2H), 5.27 (dd, J = 8.0, 5.5 Hz, 1H),4.40 (d, J = 14.6 Hz, 1H), 4.16 (d, J = 15.3 Hz, 1H), 3.89 (dd, J =22.3, 16.0 Hz, 2H), 3.44-3.35 (m, 2H), 3.19 (d, J = 6.1 Hz, 4H), 3.02(dt, J = 7.9, 4.0 Hz, 1H), 2.54 (ddd, J = 10.3, 6.7, 3.6 Hz, 1H), 2.35(dq, J = 14.0, 6.9 Hz, 1H), 2.29-2.15 (m, 1H), 1.55 (ddd, J = 10.7, 6.9,4.4 Hz, 1H), 1.42 (q, J = 7.1 Hz, 1H) 151 17 564 [M + H]⁺ (300 MHz,MeOD-d₄) δ ppm: 7.92 (s, 2H), 7.67 (dd, J = 17.5, 8.3 Hz, 4H), 7.51-7.30(m, 3H), 7.07 (s, 2H), 6.93 (q, J = 9.0, 8.0 Hz, 2H), 5.07 (s, 1H),4.51-4.23 (m, 2H), 3.94 (s, 1H), 3.67 (s, 1H), 3.36 (s, 1H), 3.16 (d, J= 18.6 Hz, 3H), 2.91 (s, 2H), 2.47 (s, 1H), 2.07 (d, J = 9.2 Hz, 1H),1.89 (s, 2H), 1.71 (d, J = 10.8 Hz, 2H), 1.12 (d, J = 32.7 Hz, 2H). 15233 [M + H]⁺ (300 MHz, CD₃OD-d₄) 8.04-7.83 (m, 8H), 7.23-7.18 (m, 628;2H), 7.06-7.00 (m, 2H), 5.27-5.22 (m, 1H), 4.42-4.35 (m, 1H), 4.19-4.12(m, 1H), 3.95-3.80 (m, 2H), 3.36-3.30 (m, 2H), 3.19-3.14 (m, 7H),3.03-2.97 (m, 1H), 2.55-2.48 (m, 1H), 2.38-2.15 (m, 2H), 1.58-1.51 (m,1H), 2.41-1.32 (m, 1H) 153 17 529 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:7.99-7.92 (m, 2H), 7.80- 7.72 (m, 2H), 7.69 (dd, J = 7.5, 1.6 Hz, 2H),7.49 (t, J = 7.6 Hz, 2H), 7.41 (t, J = 7.3 Hz, 1H), 7.11-7.02 (m, 2H),6.94 (t, J = 8.8 Hz, 2H), 5.10 (dd, J = 8.7, 5.2 Hz, 1H), 3.81 (d, J =15.7 Hz, 2H), 3.69 (s, 1H), 3.53 (d, J = 24.1 Hz, 1H), 2.79 (dp, J =11.8, 5.1 Hz, 2H), 2.60- 2.37 (m, 4H), 2.34 (s, 2H), 2.30 (dt, J = 7.7,4.1 Hz, 1H), 1.98-1.78 (m, 3H), 1.69 (q, J = 8.0 Hz, 2H), 1.06 (dt, J =9.6, 4.9 Hz, 1H), 0.98 (q, J = 6.1 Hz, 1H) 155 2 457 [M + H]⁺ (300 MHz,MeOD-d₄) δ ppm: 7.96-7.93(m, 2H), 7.24- 7.17(m, 4H), 7.04-7.00(m, 2H),5.19-5.15(m, 1H), 3.31(m, 10H), 2.98-2.92(m, 4H), 2.52-2.50(m, 1H),2.30- 2.20(m, 1H), 2.19-2.17(m, 1H), 1.55-1.51(m, 1H), 1.49- 1.35(m, 1H)156 18 [M + H]+554 (300 MHz, CD₃OD-d4) δ (ppm): 8.31 (d, J = 2.6 Hz,1H), 8.01-7.82 (m, 4H), 7.76-7.74(m, 1H), 7.22-7.10 (m, 2H), 7.06-6.92(m, 2H), 6.56-6.54 (m, 1H), 5.14-5.12 (m, 1H), 4.42-4.38 (m, 1H),4.32-4.24 (m, 1H), 3.92-3.88 (m, 1H), 3.75-3.65 (m, 1H), 3.29-3.08 (m,5H), 2.92-2.82 (m, 1H), 2.46-2.42 (m, 1H), 1.99-1.78 (m, 4H), 1.51-1.27(m, 2H) 157 7 519 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.36 (d, J = 2.6Hz, 1H), 8.08-7.98 (m, 2H), 7.97-7.88 (m, 2H), 7.80 (d, J = 1.7 Hz, 1H),7.30-7.16 (m, 2H), 7.11-6.98 (m, 2H), 6.60 (t, J = 2.2 Hz, 1H), 5.12(dd, J = 8.2, 5.2 Hz, 1H), 4.60 (d, J = 85.5 Hz, 2H), 3.59 (s, 3H),3.31-3.03 (m, 4H), 3.03-2.83 (m, 4H), 2.50 (ddd, J = 10.3, 6.6, 3.6 Hz,1H), 2.09-1.72 (m, 4H), 1.51 (ddd, J = 10.8, 6.9, 4.4 Hz, 1H), 1.43-1.31(m, 1H). 158 7; 520 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.61(s, 1H),8.04-7.95(q, 34(a); 4H), 7.90(s, 1H), 7.04-7.00(m, 2H), 6.99-6.86(m,2H), 34(b) 5.09-5.05(m, 1H), 3.80-3.61(m, 3H), 3.59-3.49(m, 1H),2.80-2.70(m, 2H), 2.51-2.30(m, 4H), 2.30-2.18(m, 4H), 1.87-1.80(m, 3H),1.65-1.62(m, 2H), 1.02-0.91(m, 2H) 159 2 529 [M + H]⁺ (400 MHz, MeOD-d₄)δ ppm: 7.95 (d, J = 7.8 Hz, 2H), 7.75 (d, J = 7.9 Hz, 2H), 7.72-7.66 (m,2H), 7.49 (t, J = 7.6 Hz, 2H), 7.40 (t, J = 7.4 Hz, 1H), 7.12-7.04 (m,2H), 6.96 (t, J = 8.7 Hz, 2H), 5.19 (s, 1H), 5.07 (s, 1H), 4.58-4.25 (m,2H), 4.10 (d, J = 18.4 Hz, 2H), 3.91 (s, 2H), 3.55 (s, 1H), 3.46 (s,1H), 2.88 (t, J = 6.9 Hz, 2H), 2.33 (s, 1H), 2.17-1.87 (m, 4H), 1.09(dt, J = 9.6, 4.9 Hz, 1H), 0.99 (t, J = 6.0 Hz, 1H) 160 2 515 [M + H]⁺(300 MHz, Methanol-d₄) δ 7.98-7.88 (m, 2H), 7.78-7.59 (m, 4H), 7.51-7.31(m, 3H), 7.17 (ddd, J = 8.2, 5.2, 2.6 Hz, 2H), 7.08-6.94 (m, 2H), 5.20(t, J = 6.7 Hz, 1H), 4.33-4.16 (m, 1H), 4.07 (d, J = 18.5 Hz, 1H),3.87-3.66 (m, 2H), 3.32 (m, 4H), 2.97 (dt, J = 7.9, 4.1 Hz, 1H),2.57-2.48 (m, 1H), 2.36-2.24 (m, 1H), 2.16 (dt, J = 14.0, 7.1 Hz, 1H),1.50 (dt, J = 10.9, 5.9 Hz, 1H), 1.35 (q, J = 7.0 Hz, 1H) 161 17 506[M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 7.92-7.90(m, 2H), 7.23- 7.17(m, 4H),7.04-7.00(m, 2H), 5.12-5.09(m, 1H), 4.48- 4.40(m, 1H), 4.31-4.21(m, 1H),3.92-3.83(m, 1H), 3.72- 3.67(m, 1H), 3.28-3.09(m, 6H), 2.95-2.93(m, 1H),2.44(m, 1H), 1.90-1.82(m, 4H), 1.47-1.44(m, 1H), 1.37- 1.35(m, 1H) 16235 :[M + H]⁺565 (300 MHz, MeOD-d4) δ ppm: 8.70-8.58 (m, 1H), 8.15- 8.00(m, 2H), 8.00-7.80 (m, 4H), 7.60-7.42 (m, 1H), 7.10-6.96 (m, 2H),6.96-6.80 (m, 2H), 5.15-5.00 (m, 1H), 4.60-4.20 (m, 2H), 4.03-3.82 (m,1H), 3.72-3.56 (m, 1H), 3.48-3.35 (m, 1H), 3.23-3.02 (m, 3H), 2.85-2.65(m, 2H), 2.35-2.20 (m, 1H), 2.00-1.80 (m, 3H), 1.78-1.50 (m, 2H),1.05-0.88 (m, 2H) 163 36 [M + H]+555 (300 MHz, Methanol-d₄): δ(ppm):9.06 (s, 2H), 8.09- 7.98 (m, 2H), 7.79-7.69 (m, 2H), 7.02 (dd, J = 8.7,5.4 Hz, 2H), 6.90 (t, J = 8.8 Hz, 2H), 5.08-4.96 (m, 1H), 4.48-4.35(m,2H), 3.92-3.81 (m, 1H), 3.68-3.63 (m, 1H), 3.42-3.35(m, 1H), 3.15-3.08(m, 3H), 2.75 (t, J = 7.1 Hz, 2H), 2.30-2.20 (m, 1H), 1.89-1.82 (m, 3H),1.64- 1.55 (m, 2H), 1.08-0.89 (m, 2H) 164 37 555 [M + H]⁺ (300 MHz,Methanol-d₄) δ 8.61 (d, J = 1.3 Hz, 1H), 8.11- 7.87 (m, 5H), 7.02 (ddd,J = 8.3, 5.4, 2.6 Hz, 2H), 6.97- 6.83 (m, 2H), 5.09-4.97 (m, 1H), 4.42(d, J = 18.0 Hz, 1H), 4.32 (s, 1H), 3.93 (t, J = 12.6 Hz, 1H), 3.64 (s,1H), 3.41 (d, J = 12.3 Hz, 1H), 3.11 (m, 3H), 2.75 (t, J = 7.4 Hz, 2H),2.26 (ddd, J = 7.5, 4.4, 3.3 Hz, 1H), 1.94- 1.79 (m, 3H), 1.65 (t, J =8.4 Hz, 2H), 1.08-0.87 (m, 2H) 165 18 489 [M + H]⁺ (300 MHz, CD3OD-d₄)δ(ppm): 8.62 (ddt, J = 4.8, 1.8, 1.0 Hz, 1H), 8.07 (dq, J = 7.9, 1.0 Hz,1H), 7.95 (tt, J = 7.5, 1.4 Hz, 1H), 7.56 (ddt, J = 7.4, 4.9, 1.2 Hz,1H), 7.21-7.08 (m, 2H), 6.98 (ddd, J = 8.7, 7.8, 1.2 Hz, 2H), 5.14 (dd,J = 7.6, 5.0 Hz, 1H), 4.45 (d, J = 14.8 Hz, 1H), 4.21 (d, J = 15.6 Hz,1H), 3.87 (t, J = 12.6 Hz, 1H), 3.65 (t, J = 11.5 Hz, 1H), 3.27-3.02 (m,6H), 2.89 (p, J = 4.0 Hz, 1H), 2.47-2.37 (m, 1H), 2.09-1.70 (m, 4H),1.49- 1.25 (m, 2H) 166 38 554 [M + H]⁺ (300 MHz, DMSO-d₆) δ 9.02 (s,1H), 8.48 (d, J = 7.8 Hz, 1H), 8.22 (s, 1H), 7.89-7.80 (m, 2H), 7.53 (t,J = 7.9 Hz, 2H), 7.36 (dd, J = 8.2, 6.5 Hz, 1H), 7.11-6.94 (m, 4H), 4.92(d, J = 7.4 Hz, 1H), 4.11 (s, 2H), 3.88 (s, 1H), 3.64 (s, 1H), 3.27 (s,2H), 3.15 (s, 1H), 3.02 (d, J = 10.6 Hz, 1H), 2.62 (t, J = 6.5 Hz, 2H),2.17 (s, 1H), 1.73 (dd, J = 24.5, 12.1 Hz, 3H), 1.49 (d, J = 10.2 Hz,2H), 0.90 (p, J = 6.5, 5.5 Hz, 2H) 167 2 [M + H]+554 (300 MHz,Methanol-d4) δ 8.24 (s, 1H), 8.05-7.94 (m, 2H), 7.73-7.61 (m, 3H), 7.16(s, 1H), 7.08-6.97 (m, 2H), 6.98-6.84 (m, 2H), 5.08-4.97 (m, 1H),4.45-4.28 (m, 2H), 3.95-3.87 (m, 1H), 3.64-3.55 (m, 1H), 3.40-3.32 (m,1H), 3.26-3.08 (m, 3H), 2.77 (t, J = 7.0 Hz, 2H), 2.28 (dt, J = 7.5, 3.9Hz, 1H), 1.90-1.85 (m, 3H), 1.65-1.57 (m, 2H), 1.10-0.89 (m, 2H) 168 39566 [M + H]⁺ (400 MHz, DMSO-d₆) δ ppm: 9.00-8.90 (d, 2H), 8.90- 8.80 (d,1H), 8.50-8.40 (d, 2H), 8.10-8.00 (d, 2H), 7.50 (t, 1H), 7.10-6.90 (m,4H), 5.00-4.80 (dd, 1H), 4.25-4.05 (m, 2H), 4.00-3.80 (m, 1H), 3.70-3.50(t, 1H), 3.30-3.25 (m, 2H), 3.25-3.10 (m, 1H), 3.10-2.95 (m, 1H),2.70-2.60 (m, 2H), 2.40-2.30 (m, 1H), 2.20-2.10 (m, 1H), 1.95-1.65 (m,3H), 1.65-1.40 (m, 2H), 1.00-0.80 (m, 2H) 169 18 471 [M + H]⁺ (400 MHz,MeOD-d₄) δ ppm: 7.91-7.88(m, 2H), 7.23- 7.17(m, 4H), 7.04-7.00(m, 2H),5.13-5.00(m, 1H), 4.38- 4.00(m, 2H), 3.97-3.70(m, 2H), 3.40-3.30(m, 2H),3.30- 3.19(m, 2H), 3.00-2.91(m, 1H), 2.57-2.43(m, 1H), 2.10- 1.73(m,4H), 1.60-1.28(m, 2H) 170 18 485 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm:7.88-7.85(m, 2H), 7.20- 7.13(m, 4H), 7.01-6.96(m, 2H), 5.13-4.91(m, 1H),4.45- 3.71(m, 4H), 3.51-3.35(m, 2H), 3.11-3.31(m, 2H), 3.00- 2.95(m,4H), 2.57-2.43(m, 1H), 2.10-1.63(m, 4H), 1.45- 1.40(m, 1H), 1.36-1.31(m,1H) 171 18 642 [M + H]⁺ ¹H NMR (300 MHz, MeOD-d₄) δ ppm: 8.08 (d, J =8.3 Hz, 2H), 8.04-7.92 (m, 4H), 7.85 (d, J = 8.3 Hz, 2H), 7.20 (dd, J =8.6, 5.2 Hz, 2H), 7.03 (t, J = 8.7 Hz, 2H), 5.18 (s, 1H), 4.43 (s, 2H),4.26 (s, 2H), 3.95 (s, 2H), 3.72 (s, 2H), 3.18 (s, 5H), 3.01-2.93 (m,1H), 2.45 (s, 1H), 2.07-1.76 (m, 5H), 1.54-1.33 (m, 3H). 172 18 566 [M +H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.20(s, 1H), 8.69(s, 1H), 8.57-8.56(m,1H), 8.27-8.18(m, 2H), 8.05-7.95 (m, 2H), 7.22-7.12 (m, 2H), 7.05-6.93(m, 2H), 5.22-5.10(m, 1H), 4.50-4.41(m, 1H), 4.32-4.21(m, 1H),3.99-3.86(m, 1H), 3.79-3.66(m, 1H), 3.39-3.02 (m, 6H), 3.00-2.81(m, 1H),2.50-2.41(m, 1H), 2.00-1.79(m, 4H), 1.52-1.41(m, 1H), 1.41-1.31(m, 1H)173 34 566 [M + H]⁺ (400 MHz, MeOD-d₄) δ ppm: 9.20(s, 1H), 8.69(s, 1H),8.57-8.56(m, 1H), 8.27-8.18(m, 2H), 8.05-7.95 (m, 2H), 7.22-7.12 (m,2H), 7.05-6.93 (m, 2H), 5.22-5.10(m, 1H), 4.50-4.41(m, 1H), 4.32-4.21(m,1H), 3.99-3.86(m, 1H), 3.79-3.66(m, 1H), 3.39-3.02 (m, 6H), 3.00-2.81(m,1H), 2.50-2.41(m, 1H), 2.00-1.79(m, 4H), 1.52-1.41(m, 1H), 1.41-1.31(m,1H) 174 18 [M + H]⁺513 (300 MHz, DMSO-d6) δ ppm: 8.00-7.90 (m, 2H),7.900- 7.730 (m, 2H), 7.36-7.11 (m, 2H), 7.10-6.90 (m, 2H), 5.21-5.02(m, 1H), 4.54-4.32 (m, 1H), 4.32-4.15 (m, 1H), 4.00-3.80 (m, 1H),3.78-3.52 (m, 1H), 3.31-3.30 (m, 1H), 3.25-3.19 (m, 2H), 3.19-3.05 (m,3H), 2.99-2.88 (m, 1H), 2.49-2.36 (m, 1H), 2.05-1.76 (m, 4H), 1.55-1.25(m, 2H) 175 7 519 [M + 1]⁺ (CD₃OD, ppm): 8.60 (s, 1H), 8.00-8.10 (m,2H), 7.88- 7.99 (m, 2H), 7.79-7.80 (m, 1H), 7.18-7.28 (m, 2H), 7.00-7.10(m, 2H), 6.59 (s, 1H), 5.05-5.20 (m, 1H), 4.20- 4.50 (m, 1H), 4.05-4.18(m, 1H), 3.70-4.00 (m, 2H), 3.38-3.42 (m, 1H), 3.21-3.28 (m, 2H),2.91-3.01 (m, 1H), 2.43-2.47 (m, 1H), 1.81-2.17 (m, 4H), 1.45-1.52 (m,1H), 1.35-1.44 (m, 1H) 176 7 477 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 8.31(d, J = 2.6 Hz, 1H), 8.00-7.81 (m, 4H), 7.75 (d, J = 1.7 Hz, 1H),7.22-7.11 (m, 2H), 7.07-6.94 (m, 2H), 6.55 (dd, J = 2.6, 1.8 Hz, 1H),4.65 (t, J = 6.8 Hz, 1H), 4.35 (dq, J = 39.0, 8.3 Hz, 2H), 4.13-3.91 (m,2H), 3.22 (t, J = 7.3 Hz, 2H), 2.93 (dt, J = 8.0, 4.1 Hz, 1H), 2.49-2.23(m, 3H), 1.84 (q, J = 11.1, 8.9 Hz, 4H), 1.51-1.27 (m, 2H). 177 7 [M +H]⁺506 (300 MHz, DMSO-d6)δ ppm: 9.11-8.12 (m, 3H), 8.62 (d, J = 2.7 Hz,1H), 8.11-8.01(m, 2H), 7.81 (d, J = 1.5 Hz, 1H), 7.31-7.20 (m, 2H),7.20-7.05 (m, 2H), 6.70-6.50 (m, 1H), 5.02-4.82 (m, 1H), 3.78-3.36 (m,8H), 3.22-3.03 (m, 2H), 3.00-2.87 (m, 1H), 2.46-2.32 (m, 1H),1.96-1.51(m, 4H), 1.50-1.35 (m, 1H), 1.35-1.22 (m, 1H). 178 7 583 [M +H]⁺ (400 MHz, MeOD-d₄) δ ppm: 8.37 (d, J = 2.6 Hz, 1H), 8.08-7.98 (m,2H), 7.98-7.88 (m, 2H), 7.80 (d, J = 1.8 Hz, 1H), 7.27-7.17 (m, 2H),7.11-6.99 (m, 2H), 6.60 (t, J = 2.2 Hz, 1H), 5.16 (dd, J = 8.2, 5.1 Hz,1H), 3.99 (d, J = 13.3 Hz, 1H), 3.92 (d, J = 13.3 Hz, 1H), 3.65 (t, J =10.7 Hz, 1H), 3.53 (dd, J = 12.9, 8.5 Hz, 1H), 3.39 (d, J = 3.9 Hz, 2H),3.31-3.11 (m, 4H), 2.97 (dt, J = 7.9, 4.0 Hz, 1H), 2.87 (s, 3H), 2.47(ddd, J = 10.3, 6.7, 3.7 Hz, 1H), 2.06-1.75 (m, 4H), 1.48 (ddd, J =10.8, 6.9, 4.4 Hz, 1H), 1.39 (q, J = 7.1 Hz, 1H). 179 18 458 [M + H]⁺(400 MHz, DMSO-d₆) δ ppm: 8.70-8.60 (d, 1H), 8.05- 7.90 (dd, 2H),7.35-7.20 (t, 2H), 7.05-6.95 (m, 4H), 4.92- 4.75 (m, 1H), 3.60-3.50 (m,6H), 3.50-3.40 (m, 2H), 2.70-2.60 (t, 2H), 2.45-2.25 (m, 1H), 2.20-2.10(m, 1H), 1.85-1.60 (m, 3H), 1.55-1.40 (m, 2H), 1.00-0.80 (m, 2H) 180 40553 [M + H]⁺ (300 MHz, MeOD-d₄) δ ppm: 7.92-7.90(d, J = 4.8 Hz, 2H),7.56-7.54(d, J = 4.8 Hz, 2H), 7.25-7.24(t, J = 2.1 Hz, 2H), 7.04-6.99(m,2H), 6.93-6.87(m, 2H), 6.30-6.28(t, J = 2.1 Hz, 2H), 5.03-4.99(m, 1H),4.55-4.25(m, 2H), 3.995-3.86(m, 1H), 3.72-3.58(m, 1H), 3.48-3.33(m, 1H),3.18-3.07(m, 3H), 2.76-2.72(m, 2H), 2.26-2.24(m, 1H), 1.89-1.82(m, 3H),1.65-1.63(m, 2H), 1.02-0.91(m, 2H) 181 41 531 [M + H]⁺ ¹H NMR (400 MHz,MeOD-d₄) δ ppm: 8.85(s, 2H), 8.48(d, J = 8.4 Hz, 2H), 8.05-7.90 (m, 2H),7.55-7.40 (m, 1H), 7.10-6.98 (m, 2H), 6.98-6.80 (m, 2H), 5.15-5.00 (m,1H), 3.88-3.45 (m, 4H), 2.88-2.70 (m, 2H), 2.60-2.35 (m, 4H), 2.35-2.20(m, 4H), 1.96-1.75 (m, 3H), 1.75-1.62 (m, 2H), 1.10-0.88 (m, 2H) 182 42491 ¹H NMR (300 MHz, DMSO-d₆): □ 10.9-10.7 (m, 1H), 9.77-9.39 (m, 2H),9.15 (d, J = 7.8 Hz, 1H), 8.64 (d, J = 2.8 Hz, 1H), 8.07-7.97 (m, 4H),7.81 (app s, 1H), 7.27- 7.11 (m, 4H), 6.61 (t, J = 2.1 Hz, 1H), 5.37 (t,J = 6.0 Hz, 1H), 4.44-4.30 (m, 2H), 3.51-3.45 (m, 4H), 3.02-2.92 (m,3H), 2.77 (s, 3H), 2.75-2.59 (m, 2H), 1.63-1.57 (m, 1H), 1.30-1.23 (m,2H) 183 2 583 184 32 579 185 2 545 186 43 515 [M + H]⁺ ¹H NMR (300 MHz,Methanol-d₄) δ 8.07-7.91 (m, 6H), 7.58 (m, 3H), 7.06-6.83 (m, 4H), 5.08(dd, J = 8.5, 5.3 Hz, 1H), 4.36 (m, 2H), 3.89 (m, 2H), 3.66 (s, 1H),3.39 (s, 1H), 3.10 (s, 3H), 2.81 (t, J = 6.7 Hz, 2H), 2.25 (dd, J = 7.2,3.9 Hz, 1H), 2.04-1.87 (m, 2H), 1.06-0.87 (m, 2H) 187 44 568 [M + H]⁺ ¹HNMR (300 MHz, MeOD-d₄) δ ppm: 7.98-7.92(m, 2H), 7.75-7.68(m, 4H),7.29-7.20 (m, 2H), 7.10-7.05 (m, 2H), 6.98-6.90 (m, 2H), 5.19-5.13 (m,1H), 4.52-4.32 (m, 2H), 4.00-3.91(m, 1H), 3.76-3.65 (m, 1H), 3.50-3.38(m, 1H), 3.22-3.11 (m, 3H), 2.91-2.85(m, 2H), 2.35-2.30 (m, 1H),2.16-1.86 (m, 3H), 1.09-0.93(m, 2H) 188 45 428 [M + H]⁺ ¹H NMR (400 MHz,MeOD-d₄) δ ppm: 7.93-7.89(m, 2H), 7.25-7.20(m, 4H), 7.07-7.01(m, 2H),4.70-4.61(m, 1H), 4.50-4.41(m, 1H), 4.40-4.29(m, 1H), 4.18-4.00(m, 2H),3.30-3.21(m, 2H), 3.18-2.90(m, 1H), 2.55-2.45(m, 1H), 2.38-2.20(m, 2H),2.00-1.74(m, 4H), 1.60-1.50(m, 1H), 1.41-1.35(m, 1H)

The invention is further illustrated by the following examples, whichmay not have been made yet or tested. The methods exemplified below mayalso be extrapolated to compounds disclosed herein which may not yethave not been made or tested. A variety of stereochemical arrangementsare shown below and are meant to demonstrate that all racemic, (R), (S),cis, and trans isomers are contemplated.

Salts and Polymorphs

Example 1 free base form is an unstable, amorphous and hygroscopicmaterial that deliquesces when subjected to high humidity, and degradesat ambient conditions over time. Various lots of Example 1 were analyzedby HPLC shortly after synthesis (analysis from 1-24 hourspost-synthesis) and re-tested for purity a unit of time later. Resultsare given in Table 3.

TABLE 3 Time Initial Re-Test Between Lot Purity Purity Tests 1 93% 82%49 days 2 81% 72% 16 days

In the salt and polymorph experiments below, the following counterionsform the following salts and are abbreviated as follows:2,5-dihydroxybenzoic acid—dihydroxybenzoate—DHBA; adipic acid—adipatesalt—ADA; benzenesulfonic acid—benzenesulfonate or besylate salt—BSA;benzoic acid—benzoate salt—BA; caprylic acid—caprylate—CYA; citricacid—citrate—CA; D-glucuronic acid—glucuronate—GA; ethanesulfonicacid—ethanesulfonate or esylate—ESA; fumaric acid—fumarate—FUA;galactaric acid—galactarate—GA; glycolic acid—glycolate—GLYA; hippuricacid—hippurate—HPA; hydrochloric acid—hydrochloride—HCl; ketoglutaricacid—ketoglutarate—KGA; L-ascorbic acid—ascorbate salt—ASBA; L-asparticacid—aspartate—ASP; L-glutamic acid—glutamate—GLU; L-lacticacid—lactate—LA; L-malic acid—malate—MA; L-tartaric acid—tartrate—TAR;maleic acid—maleate—MEA; malonic acid—malonate—MLNA; nicotinicacid—nicotinate—NA; orotic acid—orotate—ORA; oxalic acid—oxalate—OXA;phosphoric acid—phosphorate—PHOA; propionic acid—propionate—PROA;p-toluene sulfonic acid (monohydrate)-tosylate—pTSA; succinicacid—succinate—SUCA; sulfuric acid—sulfate—SUL or SO4; and thiocyanicacid—thiocyanate—TCA. Salts may form between counterions instoichiometric ratios, for example 1:1 (a mono-salt) or 2:1 (abis-salt).

Preparation of Salts of Example 1

Salt forms were prepared from a diverse range of solvent and techniquesincluding cooling, maturation, evaporation and anti-solvent addition.The possibility of forming mono and bis salts was also investigated fora number of the counter-ions.

Solvents.

Example 1 was treated with increasing volumes of solvent (2-propanol(IPA), acetone, methylethyl ketone, ethyl acetate, tetrahydrofuran,acetonitrile, 1,4-dioxane, 90:10 IPA:water, 90:10 THF:water,tert-butylmethyl ether (MTBE), and n-heptane) until the material fullydissolved or until a maximum of 100 vol. had been used. After eachaddition of solvent, the system was stirred at 25° C. for 10 min beforethe addition of a new aliquot of solvent. Example 1 was readily solublein 2-propanol (IPA), acetone, methylethyl ketone, ethyl acetate,tetrahydrofuran, acetonitrile, 1,4-dioxane, 90:10 IPA:water, and 90:10THF:water, dissolving readily in 10 vol. solvent. MTBE and n-heptanewere identified as anti-solvents when Example 1 was not soluble in 100vol. solvent.

Hydrochloride Salt.

Solutions of Example 1 were treated with neat HCl (37 wt % (12 M), 4.8μL). The samples were then stirred at 25° C. for 1 hour. The solutionswere cooled in the fridge (ca. 4° C.) for 24 hours. If no suspension wasobtained the samples were then matured at ambient/50° C. in 8-hourcycles for 4 days (Section 8.11). Any solids obtained were analyzed byXRPD. On addition of HCl to solutions of Example 1 in various solvents,many samples formed a precipitate, which after further stirring became agum. Analysis of these solids by XRPD found all to be amorphous. Resultsare given in Table 4.

TABLE 4 Obs. Obs. on after Obs. after addition of 1 Obs. after 1 coolingmaturation Solvent eq HCl hour in fridge for 4 days XRPD 2-Propanolprecipitate clear clear orange — solution solution solution Acetoneprecipitate gum gum gum amorphous Methylethyl precipitate gum gum stickysolid amorphous Ketone Ethyl Acetate precipitate gum gum sticky solidamorphous Tetrahydrofuran precipitate gum gum sticky solid amorphousAcetonitrile precipitate clear oil sticky solid amorphous solution1,4-Dioxane precipitate gum gum sticky solid amorphous 90:10 vapourclear clear orange — IPA:water evolved solution solution solution 90:10clear solution clear clear orange oil — THF:water solution solutiontert- cloudy white — — amorphous Butylmethyl solution precipitate Etherand undissolved solid n-Heptane no change on solid gum orange oil —addition dissolving (white gum)

First Salt Experiments.

Several salt experiments were undertaken in order to determine if astable, non-hygroscopic crystalline form of Example 1 could beidentified. Salts with varying crystallinity were isolated from HCl,phosphoric, sulfuric, L-tartaric, fumaric, p-toluenesulfonic and oxalicacid. In general, the crystallinity of the solids isolated was poor.

Salt Experiment 1.

Example 1 was dissolved in solvent (240 mg in 2.4 mL, 10 volumes) toform stock solutions and then dispensed into vials (400 μL/˜40 mg pervial). The solutions were treated with 1 equivalent of counterions (forhydrochloric acid (HCl), sulfuric acid (H₂SO₄ or “SO₄”), methanesulfonicacid (mesylate or “MSA”), phosphoric acid (H₃PO₄ or “PHOA”), andL-tartaric acid (TAR), 77 μL for 1M; for fumaric acid (FUA), 154 μL for0.5 M) using stock solutions. The solutions/suspensions were then cooleddown to 0° C. at 0.2° C./min and stirred at 0° C. overnight. Theresulting solids were analyzed by XRPD and those that were amorphous orpoorly crystalline were then matured for 5 days. If after cooling to 0°C. a clear solution or gum was obtained, the mother liquors were removedfrom the vials, split in two and anti-solvent (MTBE or heptane, 400 μl)added to each of the aliquots. After anti-solvent addition, the sampleswere matured for 5 days. After maturation, clear solutions and gums wereallowed to evaporate at ambient conditions. Suspensions were filtered,air-dried and characterized by XRPD. Any resulting crystalline solidswere characterized by high resolution XRPD, 1H NMR, HPLC and one weekstorage at 40° C. and 75% RH.

Crystalline solids were isolated from the experiments involvingsulfuric, phosphoric, L-tartaric acid in IPA. A further weaklycrystalline solid was also found from fumaric acid in ethyl acetate. Allother solids were found to be amorphous, although some of these showedpeak shifts by NMR, which is indicative of salt formation. Results aregiven in Table 5.

TABLE 5 Obs. on add'n Obs. Anti- of anti- after High-Res Ex. CounterionSolv. XRPD solv. solv. maturation XRPD XRPD NMR S1 HCl IPA — MTBE susp.gum Am — — S2 Heptane oil gum — — S3 SO₄ IPA WC — — sticky PartiallyPartially peak shifts, solid crystalline crystalline ~2.4 eq Sulf SulfIPA pattern 1 pattern 1 S4 MSA IPA — MTBE susp. gum Am — — S5 Heptanegum gum — — — S6 PHOA IPA Am — — white Partially Am no peak solidcrystalline shifts, ~0.4 PHOA eq IPA pattern 1 S7 TAR IPA Am — — whiteAm Crystalline peak shifts, solid TAR ~0.7 eq Form 1 acid, ~0.2 eq IPAS8 FUA IPA —* — — sticky Am — — solid S9 HCl THF — MTBE susp. gum — — —S10 Heptane susp. gum — — — S11 SO₄ THF Am — — white Am Am peak shifts,solid ~0.3 eq THF S12 MSA THF — MTBE susp. gum Am — — S13 Heptane gumgum — — — S14 PHOA THF Am — — white Am Am no peak solid shifts, ~4.6 eqTHF S15 TAR THF Am — — white Am Am peak shifts, solid ~1.4 eq acid, ~0.9eq THF S16 FUA THF Am — — white Am — — solid S17 HCl EtAc — MTBE sol'ngum — — — S18 Heptane sol'n gum — — — S19 SO₄ EtAc Am — — white Am Ampeak shifts, solid ~3.9 eq EtOAc S20 MSA EtAc — MTBE sol'n gum Am — —S21 Heptane gum gum — — — S22 PHOA EtAc Am — — white Am Am no peakssolid shifts, ~0.7 eq EtOAc S23 TAR EtAc Am — — white Am Am peak shifts,solid ~1 eq acid, ~0.4 eq EtOAc S24 FUA EtAc WC — — white WC WC peakshifts, solid FUA ~0.8 eq pattern 1 acid, ~0.2 eq EtOAc EtAC = EthylAcetate; Am = amorphous; WC = weakly crystalline; solv. = solvent; sol'n= solution; susp. = suspension; * = some ppt upon warming

Of the 16 solids obtained, four were found to be partially crystalline:sulfuric, phosphoric, L-tartaric acid in IPA. A further weaklycrystalline solid was also found from fumaric acid in ethyl acetate. Allother solids were found to be amorphous. All samples analyzed by XRPDwere placed into storage at 40° C. and 75% RH for one week. However,after one day all samples except the tartrate had deliquesced and weretherefore not characterized further. The tartrate was analyzed by XRPDafter one week at 40° C. and 75% RH and no change in form was observed,and it was further analyzed by TGA and DSC. The thermal analysis showedan initial weight loss in the TGA which corresponded to a broadendotherm in the DSC. These are likely due to the loss of IPA and waterfrom the sample. The purity of the sample by HPLC was 89.7% compared to93.6%1 for the input Example 1.

Experiment 2.

Example 1 was dissolved in methylethyl ketone (MEK) or acetonitrile(MeCN) (20 mg in 200 μl, 10 vol) at ambient conditions. The solutionswere then heated to 50° C., treated with 1.1 or 2.1 eq of acid andstirred at this temperature for 1 hour. The resultingsolutions/suspensions were then cooled down to 5° C. at 0.1° C./min andheld overnight. Any solids obtained were analyzed by XRPD and those thatwere amorphous or poorly crystalline were then matured between 50°C./ambient on an 8-hour cycle. Any clear solutions were left toevaporate slowly at ambient conditions. If the samples evaporated toform gums, these were dissolved in IPA (10 vol, 200 μL) and thenmatured. Any solutions with gums present were also matured between 50°C./ambient on an 8 hour cycle. Any resulting crystalline solids werecharacterized by high resolution XRPD, NMR, HPLC, IC (for inorganiccounter-ions) and one week storage at 40° C. and 75% RH. Table 4 showsthe experimental conditions tested.

TABLE 6 Vol for Vol for 1.1 eq 2.1 eq Counter-ion/Coformer Conc. Solvent(μL) (μL) Hydrochloric acid—HCl 1.0M THF 43 81 Sulfuric acid—SO₄ 1.0MTHF 43 81 p-Toluene sulfonic acid 1.0M THF 43 81 monohydrate—pTSA Oxalicacid—OXA 1.0M THF 43 81 Maleic acid—MEA 1.0M THF 43 81 Ketoglutaricacid—KGA 1.0M THF 43 81 2,5-Dihydroxybenzoic 1.0M THF 43 81 acid—DHBAFumaric acid—FUA 0.5M MeOH:THF 1:1 85 162 Galactaric acid—GA 1.0M THF 4381 L-Ascorbic acid—ASBA 0.5M THF 85 162 Benzoic acid—BA 1.0M THF 43 81Succinic acid—SUCA 1.0M THF 43 81 L-Tartaric acid—TAR 1.0M THF 43 81

The following experiments were done in MEK at 1:1 eq.

TABLE 7a Obs. on Obs. on Appearance Appearance XRPD acid cooling afterMaturation after after Ex. Counterion addition to 5° C. XRPD evap.length maturation maturation S25 HCl ppt gum — — 6 days gum — S26 SO₄gum ppt/gum Am — 6 days ppt Sulf pattern 2 S27 pTSA ppt clear — gum 8days clear — sol'n sol'n S28 OXA gum ppt Am — 6 days ppt OXA Form 1 S29MEA ppt gum — — 6 days gum — S30 KGA ppt gum — — 6 days gum — S31 DHBAclear clear — gum 8 days gum — sol'n sol'n S32 FUA ppt then gum — — 6days gum — redissolve S33 GA ppt ppt GA — 6 days ppt GA S34 ASBA ppt gum— — 6 days clear — sol'n S35 BA clear clear — gum 8 days ppt WC * sol'nsol'n S36 SUCA clear clear — gum 8 days ppt WC * sol'n sol'n S37 TARclear ppt Am — 6 days paste Am sol'n Am = amorphous; WC = WC; ppt =precipitate; sol'n = solution; * = some ppt upon warming.

The following experiments were done in MEK at 2:1 eq.

TABLE 7b Obs. on Obs. on Appearance Appearance XRPD acid cooling afterMaturation after after Ex. Counterion addition to 5° C. XRPD evap.length maturation maturation S38 HCl ppt gum — — 6 days gum — S39 SO₄gum ppt/gum Am — 6 days gum — S40 pTSA ppt ppt pTSA — — — — Form 1 S41OXA gum ppt Am — 6 days paste deliquesced on frit S42 MEA ppt gum — — 6days gum — S43 KGA gum gum — — 6 days gum — S44 DHBA clear clear — gum 8days gum — sol'n sol'n S45 FUA ppt then clear — gum 8 days gum —redissolve sol'n S46 GA ppt ppt GA — 6 days paste Galactaric acid S47ASBA ppt cloudy paste- — 6 days clear — sol'n WC sol'n and and red pinkgum gum S48 BA clear clear — gum 8 days ppt WC * sol'n sol'n S49 SUCAclear clear — gum 8 days clear — sol'n sol'n sol'n S50 TAR clear ppt Am— 6 days paste WC sol'n Am = amorphous; WC = WC; ppt = precipitate;sol'n = solution; * = some ppt upon warming.

The following experiments were done in MeCN at 1:1 eq.

TABLE 7c Obs. on Obs. on Appearance Appearance XRPD acid cooling afterMaturation after after Ex. Counterion addition to 5° C. XRPD evap.length maturation maturation S51 HCl ppt gum — — 4 days gum — S52 SO₄gum ppt WC — 4 days ppt Sulf pattern 3 S53 pTSA clear clear — gum 8 daysclear — sol'n sol'n sol'n S54 OXA gum ppt/gum WC — 4 days ppt OXA Form 1S55 MEA clear clear — gum 8 days gum — sol'n sol'n S56 KGA ppt gum — — 4days gum — S57 DHBA cloudy gum — — 4 days clear — sol'n sol'n S58 FUAppt gum — — 4 days gum — S59 GA ppt ppt GA — 4 days paste GA S60 ASBAcloudy red gum — — 4 days red — sol'n sol'n and gum S61 BA clear clear —gum 8 days clear — sol'n sol'n sol'n S62 SUCA clear clear — gum 8 daysppt WC * sol'n sol'n S63 TAR gum gum — — 4 days paste Am Am = amorphous;WC = WC; ppt = precipitate; sol'n = solution; * = some ppt upon warming.

The following experiments were done in MeCN at 2:1 eq.

TABLE 7d Obs. on Obs. on Appearance Appearance XRPD acid cooling afterMaturation after after Ex. Counterion addition to 5° C. XRPD evap.length maturation maturation S64 HCl ppt gum — — 4 days gum — S65 SO₄gum ppt/gum Am — 4 days gum — S66 pTSA clear ppt pTSA — sol'n Form 2 S67OXA gum ppt Am — 4 days paste Am S68 MEA clear oil — — 4 days clear —sol'n sol'n S69 KGA ppt gum — — 4 days gum — S70 DHBA ppt gum — — 4 daysclear — sol'n S71 FUA ppt gum — — 4 days gum — S72 GA ppt ppt GA — 4days paste GA S73 ASBA cloudy red 4 days red — sol'n sol'n sol'n and gumS74 BA clear clear — gum 8 days ppt WC * sol'n sol'n S75 SUCA clearclear — gum 8 days clear — sol'n sol'n sol'n S76 TAR gum gum/ Am — 4days paste Am paste Am = amorphous; WC = WC; ppt = precipitate; sol'n =solution; * = some ppt upon warming.

In total 10 solids were obtained from cooling in MEK. One of these using2.1 eq p-toluene sulphonic acid monohydrate was found to be crystalline(pTSA Form 1) and another from ascorbic acid was a paste that was weaklycrystalline. The solid obtained from galactaric acid was consistent withthe reference diffractogram for this counter-ion. All other solids fromcooling were amorphous. After maturation, crystalline solids were foundin samples from 1.1 eq additions of sulphuric acid (Sulf pattern 1) andoxalic acid (OXA Form 1).

After cooling seven solids were isolated from MeCN and analyzed by XRPD.The solid from 2.1 eq of toluene sulphonic acid monohydrate was found tobe crystalline and displayed a different XRPD diffractogram (pTSA Form2) to the reference pattern for the acid and the other tosylate solidobtained from the MEK experiment (pTSA Pattern 1). Samples from 1.1equivalent addition of sulphuric acid and oxalic acid were found to beweakly crystalline, with crystallinity improving after maturation. Thediffractogram for the sulphate (Sulf Pattern 3) was different to thosepreviously observed in the initial experiment and in MEK (Sulf Pattern 1and 2 respectively). All other solids obtained after cooling andmaturation were amorphous.

Clear solutions that were obtained on cooling in MEK and MeCN were leftto evaporate and resulted in gums. These were re-dissolved in IPA andmatured. After maturation, some of the samples yielded solids. However,all of these solids had the same XRPD pattern, despite having differentcounter-ions (benzoic and succinic acid) and acid equivalents added. ¹HNMR confirmed all of these samples had degraded to the same crystallineproduct, with trace amounts of the counter-ion present. The degradationof these samples is probably caused by the amount of further treatment(at elevated temperature) these samples underwent after the initialcooling. If salt formation was incomplete, the presence of amorphousfree base, which is known to be unstable, may also have contributed tothe degradation to an unknown crystalline compound.

Example 1 and S36 (succinate from MEK at 1:1 eq) were characterized by arange of 1D and 2D NMR experiments to further understand thisdegradation. Due to the small amount of S36 isolated, the ¹³C NMRspectra could not be fully assigned. It was clear that the phenyl ringwith the fluoride has been lost. The main degradation product is mostlikely the primary amine after loss of this ring.

Characterization of Salts.

Crystalline and partially crystalline salts were characterized furtherusing a range of techniques.

Tosylates.

Initial HPLC analysis on both tosylate salts indicated that saltformation has increased the purity to 88.0% and 90.6% compared toExample 1 which had a purity of 81.4%. Of the two forms found, pTSA Form2 is the more stable. The other salt, pTSA Form 1, becomes amorphousupon isolation and also converts to pTSA Form 2 after storage at 40° C.and 75% RH. Results are shown below in Table 8.

TABLE 8 S40 S66 Salt Tosylate from MEK 2:1 Tosylate from MeCN 2:1High-Res XRPD pTSA Form 1, pTSA Form 2 Amorphous, after isolating solid¹H-NMR (DMSO-_(d6)) Peak shifts to free base Peak shifts to free base ~2eq acid ~2 eq acid HPLC (Purity % 88.0* 90.6* AUC) Storage at 40° C.Change to pTSA Form 2 No change: pTSA and 75% RH Form 2 *= tosylate peaknot integrated

Sulfates.

Both sulfates were found to have different XRPD diffractograms (Sulfpattern 2 and 3) which were not consistent with the sulfate obtained inthe initial salt experiment which deliquesced on storage at 40° C. and75% RH (Sulf pattern 1). After storage at 40° C. and 75% RH, both solidsdisplayed the same diffractogram, Sulf pattern 2. Results are shownbelow in Table 9.

TABLE 9 S26 S52 Salt Sulfate from MEK 1:1 Sulfate from MeCN 1:1 HighResolution Weakly crystalline Weakly crystalline, XRPD Sulf pattern 2,possible Sulf pattern 3 reduction in crystallinity after isolating solid¹H-NMR (DMSO-_(d6)) Peak shifts to free base Peak shifts to free baseHPLC (Purity %, 81.4 84.0 AUC) Storage at 40° C. No change in formChange in form and 75% RH Sulf pattern 2 Sulf pattern 2 Increasedcrystallinity IC 1:0.99 1:0.98

Oxalates.

Both oxalates display the same XRPD diffractogram (OXA pattern 1),although as with the other salts, the solid from MEK loses somecrystallinity on isolation. After storage at 40° C. and 75% RH, bothoxalates changed to the same form (OXA Form 2). Results are shown belowin Table 10.

TABLE 10 Technique S28 S54 Salt Oxalate from MEK 1:1 Oxalate from MeCN1:1 High Resolution Weakly crystalline OXA OXA Form 1 XRPD Form 1,possible reduction in crystallinity after isolating solid 1H-NMR Peakshifts to free base Peak shifts to free base (DMSO-d6) ~0.2 eq MEK ~0.3eq MeCN HPLC (Purity %, 84.9 85.5 AUC) Storage at 40° C. Change in formChange in form and 75% RH OXA Form 2 OXA Form 2 Thermal analysis of DSC:Broad endotherm TGA: 5 wt % loss 30- sample from storage onset 28° C.140° C.* at 40° C. and (183.6 J/g)* 75% RH (OXA Form 2) IC 1:1.04 1:0.97*= Owing to sample availability DSC and TGA could not be performed onboth samples

Scale-Up.

The three most crystalline salt forms, the mono tartrate (TAR Form 1),mono oxalate (OXA Form 1), and bis tosylate (pTSA Form 2) were selectedfor scale up and further characterisation. This included a preliminarypolymorphism assessment.

Tartrates.

TAR Form 1 was not obtained on scale up. Instead, a new solvated form,TAR Form 2, was isolated. This form was found to be unstable to humiditywith a loss of crystallinity, conversion to a new form (TAR Form 3) anda significant drop in purity observed.

In the first experiment, Example 1 was dissolved in IPA (100 mg in 1 mL,10 vol) at ambient conditions. The solution was then heated to 50° C.,treated with 1.1 eq of L-tartaric acid (1 M in THF, 212 μL) and stirredat 50° C. for 1 hour. The resulting suspension was then cooled down to5° C. at 0.1° C./min overnight. XRPD showed only amorphous solids whichwere then matured between 50° C./ambient on an 8-hour cycle for 7 days.¹H NMR (DMSO-d₆) showed only Peak shifts to free base (˜0.5 eq IPA). Acrystalline tartrate was not obtained in this experiment even afterfurther maturation. Analysis of the amorphous tartrate by DSC indicatedno evidence of crystallization.

In the second experiment, Example 1 was dissolved in IPA (10 vol., 500mg in 5 mL) at ambient conditions. The solution was then stirred at 25°C. and treated with 1.1 eq of L-tartaric acid (1030 μL, 1 M in THF). Thesample was then stirred for 1 hour at 25° C., cooled to 0° C. at 0.2°C./min and left at 0° C. overnight. Any solids obtained were analyzed byXRPD and then matured between 50° C./ambient on an 8 hour cycle for 2days. Aliquots of the sample were taken and analyzed by XRPD duringmaturation to monitor crystallinity. The sample remained low incrystallinity and was seeded with crystalline tartrate salt (S7) after 2days. The seeded sample was returned to maturation for a further 4 daysbefore the bulk sample was filtered and dried under vacuum at ambientconditions overnight. Any resulting crystalline solids werecharacterized.

The tartrate was found to be weakly crystalline after cooling, with noimprovement seen after 2 days maturation. The sample was seeded to aidthe formation of crystalline material but after a further 4 daysmaturation XRPD analysis showed the sample to be amorphous. Afterisolation and drying, high-resolution XRPD showed the sample to havecrystallised to a new form, TAR Form 2.

The tartrate salt contained ˜1 eq of IPA by ¹H NMR, and this coupledwith the new XRPD (TAR Form 2) diffractogram confirmed a different form,most likely solvated, had been obtained. Upon heating to a point afterthe first weight loss in the TGA (75° C.) and analysing by XRPD, amixture of TAR Form 1 and 2 was observed. ¹H NMR of this material showeda decrease in the amount of IPA present. VT-XRPD indicated that thesample changed again at 120° C., which corresponds to the end of thebroad endotherm in the DSC. Other complex thermal behaviour was noted inan exotherm before degradation. GVS showed the tartrate to be veryhygroscopic taking up ˜30% w/w water, with decreased crystallinity and achange in form noted by XPPD post GVS. Both storage conditions resultedin changes in crystallinity and form but storage at 40° C. and 75% RHalso caused a significant loss in purity (>30%). Results from thisexperiment are shown below in Table 12.

Tosylates.

The bis tosylate, pTSA Form 2, was successfully scaled up and displayedthe best solid state properties of all the salts investigated. This formis believed to be a channel hydrate as water can be taken up and lostfrom the structure without changing the crystalline form. The only othercrystalline form of the tosylate observed during the study, pTSA Form 1,converted to pTSA Form 2 on storage at elevated temperature andhumidity. Limited stability studies on the tosylate indicated it hadimproved chemical stability over the free form. The bis-tosylate salthad higher purity than the corresponding free base.

In the first experiment, Example 1 was dissolved in MEK or MeCN (100 mgin 1 mL, 10 vol) at ambient conditions. The solutions were then heatedto 50° C., treated with 2.1 eq of p-toluene sulfonic acid monohydrate(1M in THF, 405 μL) and stirred constant temperature for 1 hour. Theresulting suspensions were then cooled down to 5° C. at 0.1° C./minovernight. Any solids obtained were analyzed by XRPD and those that wereamorphous or weakly crystalline were then matured between 50° C./ambienton an 8-hour cycle for 7 days. Any resulting crystalline solids werecharacterized by high resolution XRPD, ¹H NMR, HPLC, TGA, DSC and oneweek storage at 40° C. and 75% RH. After cooling the tosylates, thesolids displayed the same XRPD diffractograms, pTSA Forms 1 and 2, asobserved previously. However, after further maturation to try andimprove the crystallinity, both tosylate salts were consistent with pTSAForm 2 by XRPD. This salt was found to be a bis tosylate with higherpurity than the free base in both instances, although the solid fromMeCN was notably better. Results are shown below in Table 11.

TABLE 11 Salt Tosylate from MEK Tosylate from MeCN High Resolution pTSAForm 2 pTSA Form 2 XRPD 1H-NMR Peak shifts to free Peak shifts to free(DMSO-d6) base ~2 eq acid base ~2 eq acid HPLC (Purity %, 84.5 93.9 AUC)Storage at 40° C. No change in form No change in form and 75% RH pTSAForm 2 pTSA Form 2 DSC Broad endotherm Broad endotherm onset 28° C.(98.0 J/g) onset 28° C. (100.1 J/g) Endotherm onset 155° C. Endothermonset (12.3 J/g) followed 168° C. (38.1 J/g) by exotherm followed byexotherm TGA 2.3 wt % loss 30-60° C. 3 wt % loss 30-70° C. VT-XRPD — Nochange in form of sample up to 170° C.

The first weight loss in the TGA of the tosylate salt corresponds to abroad endotherm in the DSC. Based on the ¹H NMR, which showed nosignificant amounts of residual solvent, this suggested that pTSA Form2, is a hydrated form. Variable temperature XRPD (VT-XRPD) of tosylateshowed no change to the crystalline form of the material up to 170° C.,after which the sample melts. This indicates that pTSA Form 2 could be achannel hydrate in which the water can move in an out of the structurewithout affecting the crystalline lattice.

In the second experiment, Example 1 was dissolved in acetonitrile (10vol., 500 mg in 5 mL) at ambient conditions. The solution was thenstirred at 50° C. and treated with 2.1 eq of p-toluenesulfonic acidmonohydrate (2060 μL, 1 M in THF). The sample was then stirred for 1hour at 50° C., cooled to 5° C. at 0.1° C./min and left at 5° C.overnight. Any solids obtained were analyzed by XRPD and then maturedbetween 50° C./ambient on an 8 hour cycle for 1 day. Aliquots of thesample were taken and analyzed by XRPD during maturation to monitorcrystallinity. No notable change in crystallinity was observed betweenthe XRPD diffractograms pre and post maturation. The bulk sample wasfiltered and dried under vacuum at ambient conditions overnight. Anyresulting crystalline solids were characterized. Although the tosylatewas crystalline after cooling, it was matured to see if any increase incrystallinity would be obtained. The sample was removed from maturationafter 1 day when no change to the sample was seen by XRPD.

The XRPD diffractogram of the tosylate was found to be consistent withpTSA Form 2, as expected. Weight loss in the TGA corresponding to thebroad endotherm in the DSC, indicated this is a hydrated form. This isfurther supported by the ₁H NMR which showed no significant amounts ofresidual solvent. The sample displays a melt, endotherm (onset 170° C.)in the DSC. GVS of the material shows the sample to have a fullyreversible uptake and loss of water, only taking up ˜5% w/w water over afull cycle. No change in form was observed. Storage at both 40° C. and75% RH and 25° C. and 95% RH showed no changes in the form or purity ofthe sample. Results from this experiment are shown below in Table 12.

Oxalates.

A new form of the oxalate was also obtained on scale up. This material,named OXA Form 3, was very similar to OXA Form 1 isolated in the initialexperiment. OXA Form 3 is believed to be a hydrate form that readilyconverts to a higher hydrate on exposure to humidity. This higherhydrate, OXA Form 2, is hygroscopic but stable to elevated humidity.This form was also obtained by maturation of OXA Form 3 in THF:Water(95:5) in the polymorphism assessment.

Example 1 was dissolved in acetonitrile (10 vol., 500 mg in 5 mL) atambient conditions. The solution was then stirred at 50° C. and treatedwith 1.1 eq of oxalic acid (1030 μL, 1 M in THF). The sample was thenstirred for 1 hour at 50° C., cooled to 5° C. at 0.1° C./min and left at5° C. overnight. Any solids obtained were analyzed by XRPD and thenmatured between 50° C./ambient on an 8 hour cycle for 2 days. Aliquotsof the sample were taken and analyzed by XRPD during maturation tomonitor crystallinity. No notable change in crystallinity was observedbetween the XRPD diffractograms pre and post maturation. The bulk samplewas filtered and dried under vacuum at ambient conditions overnight. Anyresulting crystalline solids were characterized. Although the oxalatewas crystalline after cooling, it was matured to see if any increase incrystallinity would be obtained. The samples were removed frommaturation after 2 days, when no change to the sample was seen by XRPD.

The oxalate was found to have an XRPD diffractogram that was similar toOXA Form 1 with some differences. OXA Form 3 it may be a morecrystalline sample of OXA Form 1. The broad endotherm in the DSC andfirst weight loss in the TGA, suggest the oxalate is a hydrated form.The form of oxalate was found to be unchanged by temperatures up to 140°C., suggesting the water is in channels and not bound within the crystalstructure. As with the tartrate, the oxalate displayed an exothermbefore degradation in the DSC. Although the form of oxalate is notchanged by temperatures, it is by humidity. Storage at both 40° C. and75% RH and 25° C. and 95% RH changed the form of the material to OXAForm 2, with no affect on the purity of the samples. OXA Form 2 islikely to be a higher hydrated form. GVS shows the sample to behygroscopic taking up ˜12% w/w water during a full cycle, post GVS XRPDshows a change in form to OXA Form 2 post GVS. Results from thisexperiment are shown below in Table 12.

Results from the larger scale-up experiments above are shown below inTable 12.

TABLE 12 TAR Form 2 High Res (Amorphous before XRPD isolation) pTSA Form2 OXA Form 3* ¹H-NMR Peaks shifts to free Peak shifts to free base Peakshifts to free base (DMSO-d6) base ~2 eq acid trace MeCN ~1 eq acid ~1eq IPA HPLC (Purity 97.3 97.5 97.7 %, AUC) PLM Agglomerated particlesLath shaped particles Agglomerated particles Small amount ofBirefringence Birefringence birefringence DSC Broad endotherm onsetBroad endotherm onset Broad endotherm onset 46° C. 29° C. 34° C. (155.5J/g) (97.9 J/g) (117.1 J/g) Endotherm onset 124° C. Endotherm onset 170°C. Endotherm onset 115° C. (3.8 J/g) followed by (46.3 J/g) (1.1 J/g)exotherm onset 130° C. Exotherm onset 155° C. (70.2 J/g) (30.7 J/g) TGA4.4 wt % loss 30-70° C. 2.9 wt % loss 40-80° C. 4.4 wt % loss 40-80° C.1.5 wt % loss 110-130° C. XRPD after Isotherm at 75° C. Isotherm at 100°C. Isotherm at 100° C. first TGA Mixture of forms No change in form Nochange in form weight loss TAR Form 1 and 2 NMR: ~0.15 eq IPA KarlFischer 7.0% 4.6% 7.5% GVS Very hygroscopic Fully reversible HygroscopicReversible after 1st ~5% w/w water uptake Reversible after 1st uptakebetween uptake ~30% w/w water 0 and 90% RH ~4% ~12% w/w water uptakebetween 0 and w/w water uptake uptake between 90% RH between 0 and 90%RH High res XRPD: 0 and 30% RH High res XRPD: Change in form High resXRPD: Change in form TAR Form 3 No change in form OXA Form 2 Decreasedcrystallinity pTSA Form 2 Storage @ 40° C./ Change in formTAR No changein form Change in form 75% RH Form 3 pTSA Form 2 OXA Form 2 Decreasedcrystallinity Purity: 97.4% Purity: 97.8% Purity: 64.2% Storage @ 25°C./ Change in form TAR No change in form Change in form 95% RH Form 3pTSA Form 2 OXA Form 2 Decreased crystallinity Purity: 97.5% Purity:97.9% Purity: 95.7% VT-XRPD Change in form at 120° C. No change in formon No change in form of TAR pattern 4 heating♦. sample up to 140° C.Sample melted at 140° C. IC 1:1.00 1:2.1 1:1.03 Yield ~580 mg, ~92% ~620mg, ~93% ~500 mg, ~86% Kinetic >23.2 mg/ml at pH 2.1 16.5 mg/ml at pH1.3 >25.6 mg/ml at pH 1.6 Solubility SGF 2 hours *= Similar to OXApattern1, ♦= Analysis performed on earlier batch

Amorphous Example 1 (10 mg) was treated with increasing volumes ofsolvent at 25° C. with stirring (Table #). Clear solutions were observedin most cases. For those instances where dissolution was not observed,the samples were placed at 50° C. for 10 minutes. To all samples stillin suspension, a further 10 volumes of solvents were added and thesample placed at 25° C. The temperature was then once again increased to50° C. and maintained until the end of the experiment. Further solventwas added until the material fully dissolved or until a maximum of 70volumes had been used. Hydrochloric acid was then added (1.1equivalents) to all samples. A cooling ramp was set to 5° C. at 0.1°C./min and the solutions or oils were stirred at this temperatureovernight, yielding oils or solutions.

Solutions were evaporated to dryness (ambient conditions), or treatedwith anti-solvent (TBME, 15 μL, stirring at 25° C.). Any oils or gumswere subjected to sonication, followed by maturation between ambient and50° C. for ca. 4 hours at each temperature (Section 7.7). Oils weresubsequently treated with anti-solvent and matured further at the aboveconditions. Results are shown below in Table 13.

TABLE 13 10 10 20 20 40 70 vol vol vol vol vol vol 25° 50° 25° 50° 50°50° Add'n At Further Further Solvent C. C. C. C. C. C. of HCl 5° C. workResults work Results Diethyl oil oil oil oil oil oil oil + oil SON + oilAS + oil ether turbid MAT MAT Dimethyl # sol'n sol'n Evap. sol'n AS +oil sulfoxide (glass STIR slide) sol'n Propyl # turbid oil SON + oilAS + oil acetate MAT MAT Methyl # turbid oil SON + oil AS + oil acetateMAT MAT Isopropyl # turbid oil SON + oil AS + oil acetate MAT MAT MIBK #turbid oil SON + oil AS + oil MAT MAT MEK # turbid oil SON + oil AS +oil MAT MAT 1-Butanol # sol'n sol'n Evap. oil + AS + oil sol'n STIR2-Propanol # sol'n sol'n AS + oil + AS + oil STIR sol'n STIR 1-Propanol# sol'n sol'n Slow oil AS + oil Evap. STIR Water x x x oil oil oil oil,sol'n Evap. sol'n Evap. oil turned (glass into slide) sol'n Acetone #turbid oil SON + oil AS + oil MAT MAT Ethanol # sol'n sol'n AS + oil +AS + oil STIR sol'n STIR Anisole # turbid oil SON + oil AS + oil MAT+MAT Methanol # sol'n sol'n AS + oil + AS + oil STIR sol'n STIRCyclohexane oil oil oil oil oil oil oil + oil SON + oil AS + oil ppt MATMAT Toluene # turbid oil SON + oil AS + oil MAT MAT Acetonitrile # sol'noil SON + oil AS + oil MAT MAT Dichloromethane # turbid oil SON + oilAS + oil MAT MAT IPA:Water # sol'n sol'n Slow oil AS + oil (85:15) Evap.STIR MeOH:Water # sol'n sol'n Slow oil AS + oil (85:15) Evap. STIREtOH:Water # sol'n sol'n Slow oil AS + oil (85:15) Evap. STIRAcetone:Water # sol'n sol'n Slow oil AS + oil (95:5) Evap. STIRAcetone:Water # sol'n sol'n Slow oil AS + oil (90:10) Evap. STIR # =clear solution; x = suspension; sol'n = solution; SON = sonication 30min; MAT = maturation ambient-50° C., 4 h each, 4 days; STIR = constant25° C.; AS = anti-solvent addition TBME 150 μL;

Dissolution of amorphous Example 1 was observed in most solvents at 25°C. (10 volumes). Exceptions were water, diethyl ether and cyclohexane,where oils eventually formed. No solids were obtained after the additionof hydrochloric acid. Turbidity was observed in most instances uponaddition of the acid, but oils were observed after the cooling ramp, andalso after subsequent sonication, anti-solvent addition with stirringand/or maturation.

Second Salt Experiments. Amorphous Example 1 was dissolved in1-propanol, acetone or isopropyl acetate (1380 mg in 13.8 mL, 10volumes) at 25° C. The equivalent to 30 mg Example 1 (300 μL) waspipetted into vials, which were then placed at 50° C. The selected acidswere added at 50° C. The temperature was maintained for 1 hour and acooling ramp to 5° C. at 0.1° C./min was set up overnight.

Any precipitates observed were filtered and analyzed by XRPD. Theremaining oils or solutions were subjected to anti-solvent additions.TBME (10 volumes) was used as anti-solvent for the 1-propanolexperiments. n-Heptane (10 volumes) was the anti-solvent added to theacetone experiments, and hexane (7 volumes) was added to the isopropylacetate experiment. The samples, where applicable, were further treatedby maturation, cooling or evaporation. Finally, all oils and gums weredried under vacuum at room temperature over one week. Further solvent(IPA/5% water, nitromethane or toluene, 8 volumes) was added. Furthermaturation between ambient conditions and 50° C. was set up, 4 hours ateach temperature, for a total of 11 days.

Crystalline or partially crystalline materials were isolated forsulfate, benzenesulfonate, oxalate, fumarate, L-malate, citrate andethanesulfonate salts. Oxalate salts were not characterized owing to thefact that these salts were reported in the previous salt selectionproject.

A number of different XRPD patterns were observed for the attemptedsulfate salts, including one (SUL3) observed in the experiment above.One form of benzenesulfonate salt was observed in three instances, from1-propanol and acetone. One form of fumarate salt was observed fromIPAc/TBME, independent of the amount of acid used. Another poorlycrystalline fumarate, denoted as FUA2, was observed after maturation inIPA/5% water. Finally, partially crystalline L-malate, citrate andethanesulfonate salts were observed. The results are summarised inTables 12a-12c.

Results from salt experiments performed in 1-propanol are given below inTable 14a.

TABLE 14a Obs. Maturation Obs. Obs After in IPA/ Counter- on after Obsat TBME Previous 5% Ex. ionr Eq.s add'n 1 h 5° C. add'n XRPD ProcedureProcedure water S77 HCl 1.1 sol'n sol'n sol'n oil n/a evap. oil oil S78HCl 2.2 sol'n sol'n sol'n oil n/a evap. oil oil S79 SUL 1.1 ppt ppt pptn/a SUL3* n/a n/a n/a S80 SUL 2.2 ppt ppt ppt n/a Mainly n/a n/a n/aamorphous S81 SUL 0.6 ppt ppt ppt n/a SUL4^(#) n/a n/a n/a S82 TCA 1.1turbid turbid turbid turbid n/a evap. oil oil S83 TCA 2.2 turbid turbidturbid turbid n/a evap. oil oil S84 BSA 1 sol'n sol'n sol'n pptdeliquesced n/a n/a n/a S85 BSA 2 sol'n sol'n ppt n/a BSA1 LC^(#) n/an/a n/a S86 OXA 1.1 ppt ppt ppt n/a OXA1* n/a n/a n/a S87 OXA 2.2 pptppt ppt n/a gel maturation oil oil 25-5° C. S88 OXA 0.6 ppt ppt ppt n/adeliquesced n/a n/a n/a S89 ASP 1.1 soln/ turbid/ ppt n/a insuff. evap.insuff. n/a acid acid S90 ASP 2.2 soln/ soln/ turbid ppt insuff. evap.insuff. n/a acid acid S91 ASP 0.6 soln/ soln/ turbid/ ppt evap. insuff.n/a acid acid oil insuff. S92 MEA 1.1 sol'n sol'n sol'n oil n/a evap.oil oil S93 MEA 2.2 sol'n sol'n oil oil n/a evap. oil oil S94 PHOA 1.1ppt ppt ppt n/a deliquesced n/a n/a n/a S95 PHOA 2.2 ppt ppt ppt n/aamorphous n/a n/a n/a S96 ESA 1 sol'n sol'n sol'n sol'n n/a −20° C.sol'n oil S97 ESA 2 sol'n sol'n sol'n ppt deliquesced n/a n/a n/a S98GLU 1.1 soln/ turbid/ ppt n/a insuff. evap. insuff. n/a acid acid S99GLU 2.2 soln/ soln/ turbid ppt insuff. evap. insuff. n/a acid acid S100GLU 0.6 soln/ soln/ turbid ppt n/a evap. insuff. n/a acid acid S101 MLNA1.1 sol'n sol'n oil oil n/a evap. oil oil S102 MLNA 2.2 sol'n sol'n oiloil n/a evap. oil oil S103 FUA 1.1 sol'n sol'n sol'n oil n/a evap. oiloil S104 FUA 2.2 sol'n sol'n sol'n oil n/a evap. oil FUA2 LC^(#) S105FUA 0.6 sol'n sol'n sol'n oil n/a evap. oil oil S106 CA 1.1 ppt ppt pptn/a deliquesced^(&) n/a n/a n/a S107 CA 2.2 ppt ppt ppt n/adeliquesced^(&) n/a n/a n/a S108 GA 1.1 sol'n sol'n sol'n sol'n n/a −20°C. sol'n n/a S109 GA 2.2 sol'n sol'n sol'n sol'n n/a −20° C. sol'n n/aS110 MA 1.1 ppt sol'n oil ppt insuff. evap. insuff. n/a S111 MA 2.2 pptppt gum ppt insuff. evap. insuff. n/a S112 HPA 1.1 sol'n sol'n gum oiln/a evap. insuff. n/a S113 HPA 2.2 sol'n sol'n sol'n sol'n n/a −20° C.sol'n oil S114 GLYA 1.1 sol'n sol'n sol'n sol'n n/a −20° C. sol'n oilS115 GLYA 2.2 sol'n sol'n sol'n sol'n n/a −20° C. sol'n oil S116 LA 1.1sol'n sol'n sol'n sol'n n/a −20° C. sol'n oil S117 LA 2.2 sol'n sol'nsol'n sol'n n/a −20° C. sol'n oil S118 ADA 1.1 sol'n sol'n sol'n sol'nn/a −20° C. sol'n oil S119 NA 1.1 soln/ turbid/ turbid/ sol'n n/a −20°C. sol'n oil acid acid acid S120 PROA 1.1 sol'n sol'n sol'n sol'n n/a−20° C. sol'n oil S121 CYA 1.1 sol'n sol'n sol'n sol'n n/a −20° C. sol'noil S122 ORA 1.1 soln/ turbid/ turbid/ turbid/ insuff. evap. insuff. n/aacid acid acid ppt S123 GLU 1.1 soln/ turbid/ ppt n/a insuff. evap.insuff. n/a acid acid ppt = precipitate; sol'n = solution; −20° C. =placed in freezer; acid = where acid was added as neat solid, acidparticles were observed as opposed to precipitation of further solid;^(#) = new crystalline or partially crystalline solids; insuff. =insufficient material for analysis; LC = low crystallinity; * =crystalline or partially crystalline solids previously observed; ^(&) =analyzed by XRPD although evidence of deliquescence was observed.

Results from salt experiments performed in acetone are given below inTable 14b.

TABLE14b Obs. Obs. Obs Obs After Maturation on after at TBME Previous inEx. Counterion Eq.s add'n 1 h 5° C. add'n XRPD Procedure ProcedureNitromethane S124 HCl 1.1 turbid oil oil oil n/a maturation oil oil S125HCl 2.2 ppt oil oil oil n/a maturation oil oil S126 SUL 1.1 ppt ppt pptn/a SUL5 ^(#) n/a n/a n/a S127 SUL 2.2 ppt ppt ppt n/a Deliquesced^(&)n/a n/a n/a S128 SUL 0.6 ppt ppt ppt n/a Mainly n/a n/a n/a amorphousS129 TCA 1.1 ppt ppt turbid turbid n/a maturation oil oil S130 TCA 2.2ppt ppt turbid turbid n/a maturation oil oil S131 BSA 1 sol'n sol'n pptn/a BSA1 LC^(#) n/a n/a n/a S132 BSA 2 turbid turbid ppt n/a BSA1 LC^(#)n/a n/a n/a S133 OXA 1.1 oil ppt ppt n/a OXA1* n/a n/a n/a S134 OXA 2.2oil ppt ppt n/a n/a (gel) maturation gel oil S135 OXA 0.6 ppt ppt pptn/a OXA1* n/a n/a n/a S136 ASP ### oln/acid urbid urbid turbid insuff.maturation oil turbid S137 ASP 2.2 soln/ turbid turbid turbid insuff.evap. turbid oil acid S138 ASP 0.6 soln/ turbid turbid turbid insuff.evap. turbid oil acid S139 MEA 1.1 sol'n sol'n sol'n oil n/a maturationoil oil S140 MEA 2.2 turbid turbid sol'n oil n/a maturation oil oil S141PHOA 1.1 ppt ppt ppt n/a amorphous n/a n/a n/a S142 PHOA 2.2 ppt ppt pptn/a amorphous n/a n/a n/a S143 ESA 1 sol'n sol'n sol'n oil n/amaturation oil oil S144 ESA 2 turbid turbid ppt n/a Deliquesced n/a n/an/a S145 GLU 1.1 soln/ turbid turbid turbid n/a maturation turbid oilacid S146 GLU 2.2 soln/ turbid turbid turbid n/a maturation turbid oilacid S147 GLU 0.6 soln/ turbid turbid turbid n/a maturation turbid oilacid S148 MLNA 1.1 turbid turbid oil oil n/a maturation oil oil S149MLNA 2.2 turbid oil oil oil n/a maturation oil oil S150 FUA 1.1 sol'nsol'n sol'n oil n/a maturation oil oil S151 FUA 2.2 sol'n sol'n sol'noil n/a maturation oil Amorphous solid S152 FUA 0.6 sol'n sol'n sol'noil n/a maturation oil oil S153 CA 1.1 ppt oil oil oil n/a maturationoil oil S154 CA 2.2 ppt gum gum oil n/a maturation oil oil S155 GA 1.1sol'n sol'n sol'n sol'n n/a −20° C. sol'n oil S156 GA 2.2 sol'n sol'nsol'n sol'n n/a −20° C. sol'n oil S157 MA 1.1 ppt turbid gum oil n/amaturation oil oil S158 MA 2.2 ppt turbid gum oil n/a maturation oil oilS159 HPA 1.1 sol'n sol'n sol'n oil n/a maturation oil oil S160 HPA 2.2sol'n sol'n gum sol'n n/a −20° C. sol'n oil S161 GLYA 1.1 sol'n sol'noil oil n/a maturation oil oil S162 GLYA 2.2 turbid sol'n oil oil n/amaturation oil oil S163 LA 1.1 sol'n sol'n sol'n oil n/a maturation oiloil S164 LA 2.2 sol'n sol'n sol'n oil n/a maturation oil oil S165 ADA1.1 sol'n sol'n sol'n oil n/a maturation oil oil S166 NA 1.1 soln/acidsol'n sol'n sol'n n/a −20° C. sol'n oil S167 PROA 1.1 sol'n sol'n sol'nsol'n n/a −20° C. sol'n oil S168 CYA 1.1 sol'n sol'n sol'n sol'n n/a−20° C. sol'n oil S169 ORA 1.1 soln/acid turbid turbid turbid n/amaturation turbid oil ppt = precipitate; sol'n = solution; −20° C. =placed in freezer; acid = where acid was added as neat solid, acidparticles were observed as opposed to precipitation of further solid;^(#) = new crystalline or partially crystalline solids; insuff. =insufficient material for analysis; LC = low crystallinity; * =crystalline or partially crystalline solids previously observed; ^(&) =analyzed by XRPD although evidence of deliquescence was observed.

Results from salt experiments performed in isopropyl acetate are givenbelow in Table 14c.

TABLE 14c Obs. Obs Obs Obs. After Maturation on after at TBME Prev. inEx. Counterion Eq.s add'n 1 h 5° C. add'n XRPD Procedure ProcedureToluene S170 HCl 1.1 ppt oil oil oil maturation oil n/a oil S171 HCl 2.2ppt oil oil turbid maturation turbid n/a oil S172 SUL^(a) 1.1 ppt pptppt n/a n/a n/a SUL5^(#) n/a S173 SUL^(a) 2.2 ppt ppt ppt n/a n/a n/adeliquesced^(&) n/a S174 SUL^(a) 0.6 ppt ppt ppt n/a n/a n/a SUL5^(#)n/a S175 TCA 1.1 turbid turbid oil turbid maturation oil n/a oil S176TCA 2.2 turbid turbid ppt n/a n/a n/a deliquesced n/a S177 BSA 1 pptppt/ oil turbid maturation oil n/a oil gum S178 BSA^(a) 2 ppt ppt/ pptn/a n/a n/a amorphous n/a gum S179 OXA^(a) 1.1 ppt ppt ppt n/a n/a n/an/a OXA1 LC* S180 OXA 2.2 ppt ppt ppt n/a n/a n/a Amorphous* n/a S181OXA^(a) 0.6 ppt ppt ppt n/a n/a n/a OXA1 LC* n/a S182 ASP 1.1 soln/turbid ppt n/a n/a n/a mainly n/a acid amorphous^($) S183 ASP 2.2 soln/turbid ppt n/a n/a n/a deliquesced n/a acid S184 ASP 0.6 soln/ turbidoil turbid maturation oil n/a oil acid S185 MEA 1.1 ppt oil oil turbidmaturation oil n/a oil S186 MEA 2.2 ppt oil oil turbid maturation oiln/a oil S187 PHOA^(a) 1.1 ppt ppt ppt n/a n/a n/a Amorphous n/a S188PHOA^(a) 2.2 ppt ppt ppt n/a n/a n/a Amorphous n/a S189 ESA 1 ppt gumoil turbid maturation oil n/a oil S190 ESA^(a) 2 ppt gum ppt n/a n/a n/aESA1 LC# n/a S191 GLU 1.1 soln/ turbid turbid turbid maturation oil n/aoil acid S192 GLU^(a) 2.2 soln/ turbid ppt n/a n/a n/a Glutamic n/a acidacid S193 GLU 0.6 soln/ turbid turbid turbid maturation oil n/a oil acidS194 MLNA 1.1 ppt gum gum turbid maturation oil n/a oil S195 MLNA 2.2ppt gum gum turbid maturation oil n/a oil S196 FUA^(a) 1.1 ppt gum gumturbid maturation ppt FUA1 n/a S197 FUA 2.2 sol'n sol'n sol'n turbidmaturation oil n/a amorphous solid S198 FUA^(a) 0.6 ppt ppt ppt n/a n/an/a FUA1 n/a S199 CA 1.1 ppt ppt ppt n/a n/a n/a CA1 LC^(@) n/a S200CA^(a) 2.2 ppt ppt ppt n/a n/a n/a amorphous n/a S201 GA 1.1 sol'n sol'noil turbid maturation oil n/a oil S202 GA 2.2 sol'n turbid oil turbidmaturation oil n/a oil S203 MA 1.1 ppt ppt ppt n/a n/a n/a deliquescedn/a S204 MA^(a) 2.2 ppt ppt gum turbid maturation ppt MA1 LC n/a S205HPA 1.1 turbid turbid sol'n turbid maturation oil n/a oil S206 HPA 2.2turbid turbid sol'n turbid maturation oil n/a oil S207 GLYA 1.1 ppt pptgum turbid maturation oil n/a oil S208 GLYA 2.2 ppt gum gum turbidmaturation oil n/a oil S209 LA 1.1 sol'n sol'n sol'n turbid maturationoil n/a oil S210 LA 2.2 sol'n sol'n sol'n turbid maturation oil n/a oilS211 ADA 1.1 oil oil gum turbid maturation sol'n n/a oil S212 NA 1.1soln/ sol'n sol'n turbid maturation sol'n n/a oil acid S213 PROA 1.1sol'n sol'n sol'n sol'n maturation oil n/a oil S214 CYA 1.1 sol'n sol'nsol'n sol'n maturation oil n/a Solid- impurity S215 ORA 1.1 soln/ pptppt n/a n/a n/a Deliquesced^(&,$) n/a acid ppt = precipitate; sol'n =solution; −20° C. = placed in freezer; acid = where acid was added asneat solid, acid particles were observed as opposed to precipitation offurther solid; ^(#) = new crystalline or partially crystalline solids;insuff. = insufficient material for analysis; LC = low crystallinity; *= crystalline or partially crystalline solids previously observed; ^(&)= analyzed by XRPD although evidence of deliquescence was observed; ^($)= small peaks consistent with L-aspartic acid; ^(a) = vacuum driedsamples; ^(@) = isolated by drying only (insufficient solid to filter).

The fumarate salt obtained after maturation from IPA/5% water (FUA2) waspoorly crystalline. ¹H NMR analysis was carried out suggesting thatalthough salt formation had occurred (chemical shifts consistent withother salts), an excess of fumaric acid was present.

The poorly crystalline solid obtained from caprylic acid in IPAc wasconsistent with a crystalline impurity identified during the priorexperiments. The ¹H NMR spectrum is not consistent with the other saltsformed throughout this project and suggests degradation (the integralsare different and there are a number of extra peaks not attributable toeither the compound or caprylic acid).

Basic characterisation of the sulfate, benzenesulfonate, fumarate,ethanesulfonate, L-malate, and citrate salts was carried out whereenough material was available. In the instances where insufficientmaterial was available for characterisation, preparation was repeated ona slightly larger scale.

Characterization of Salts.

Crystalline or partially crystalline novel salts obtained throughoutthis study were characterized further.

Amorphous Example 1 was dissolved in 1-propanol or acetone (60 mg in 0.6mL, 10 volumes) at 50° C. The selected acids (see table below fordetails) were added at 50° C. Precipitates were observed from bothsulfate salt formation attempts at 50° C. Anti-solvent was added at 50°C. to the benzenesulfonate salt attempt, where precipitation wasobserved (TBME, 0.6 mL, 10 volumes). The temperature was maintained for1 hour and a cooling ramp to 5° C. at 0.1° C./min was set up overnight.Solids were filtered, dried under vacuum overnight at ambient conditionand analyzed by XRPD. Results are shown below in Table 15.

TABLE 15 Eqs Target Form Acid added Solvent Form Obtained Sulfuric 1.11-propanol SUL5 SUL3 Benzenesulfonic 2.2 1-propanol/TBME BSA1 BSA1Sulfuric 0.6 acetone SUL4 SUL5

Crystalline or partially crystalline salts obtained throughout thisstudy were 257 characterized further, e.g. for stoichiometry, solventcontent, chemical purity by HPLC and stability at elevated temperatureand humidity.

Sulfates.

SUL3 and SUL5 are both crystalline and confirmed by ion chromatographyas mono salts. Both materials remained as solids after storage at 40°C./75% RH for four days, although changes were observed by XRPD (SUL3showed changes whereas SUL5 became amorphous). Both materials displayeda weight loss (6.2% below 190° C. and 4.4% below 175° C. respectively)on heating in thermogravimetric experiments. SUL3 also contained roughlyone mole equivalent of 1-propanol by ¹H NMR indicating it may be asolvated form. HPLC analysis of this form showed a purity of 98.8%.

Benzenesulfonates.

BSA1 is partially crystalline and was confirmed as abis-benzenesulfonate salt by ¹H NMR. DSC showed a broad endotherm,consistent with a TG weight loss of 5.1% below 150° C., followed by asharper endotherm with onset at 115.5° C. This material deliquesced uponstorage at 40° C./75% RH overnight, and had a purity of 97.8%.

Fumarates.

FUA1 is poorly crystalline and was confirmed as a mono-fumarate salt by¹H NMR. A TG weight loss of 3.6% w/w was also observed below 150° C. DSCanalysis of the sample was complex with multiple unresolved events. Thismaterial has a purity of 94.3% and deliquesced upon storage at 40°C./75% RH overnight.

The partially crystalline ESA1 is confirmed as a bis-ethanesulfonatesalt. This material deliquesced upon storage at 40° C./75% RH overnight.A weight loss of 2.8% w/w was observed below 150° C.

The partially crystalline MA1 was also 257 characterized. Thestoichiometry could not be accurately determined due to overlappingpeaks in the ¹H NMR spectrum. This material has a purity of 94.7% anddeliquesced upon storage at 40° C./75% RH overnight. A TG weight loss of7.4% w/w was observed below 150° C.

Finally, the partially crystalline CA1 was also characterized. Thepurity was the lowest of all isolated salts, with a value of 87.8%. Thematerial also deliquesced upon storage at 40° C./75% RH overnight andthe stoichiometry could not be accurately determined due to overlappingpeaks in the ¹H NMR spectrum. Results are shown below in Table 16.

TABLE 16 Storage Salt/ Stoichiometry 40° C./ Purity by Solvent XRPD(¹HNMR or IC) 75% RH TGA DSC HPLC (¹HNMR) SUL3 Mono salt Change 6.2%Multiple 98.80% 1 equivalent (0.9 equivalents) observed weight lossoverlapping events 1-ProH after 4 days below 190° C. SUL5* Mono saltMainly 4.4% Multiple Insufficient 0.1 (0.9 equivalents) amorphous weightloss overlapping events material equivalents solid after 4 below acetonedays 170° C. BSA1 Bis salt Deliquesced 5.1% Solvent loss broad 97.80%0.6 (1.8 equivalents) overnight weight loss endotherm (onset equivalentsbelow 34.8° C., 1-ProH 150° C. ΔH = 25.8 J/g) followed by sharperendotherm (onset 115.5° C., ΔH = 17.0 J/g) FUA1* Mono salt Deliquesced3.6% Multiple 94.30% 0.1 (1.1 equivalents) overnight weight lossoverlapping events equivalents below IPAc 150° C. ESA1 Bis saltDeliquesced 2.8% Multiple Insufficient No (2.0 equivalents) overnightweight loss overlapping events material significant below residual 150°C. solvent MA1 Unable to Deliquesced 7.4% Multiple 94.70% 0.3 eqs IPAc,LC** quantify, peak overnight weight loss overlapping events 1.5 eqs THFoverlapping below 150° C. CA1 Unclear, peak Deliquesced InsufficientMultiple 87.80% 0.4 eqs IPAc overlapping overnight material overlappingevents LC = low crystallinity; *Mono salts were obtained although hemior bis salts were targeted; **sample went amorphous at RTPreparation of Polymorphs of Salts of Example 1

A preliminary polymorphism assessment was carried out on the three mostcrystalline salt forms, the mono tartrate (TAR Form 1), mono oxalate(OXA Form 1), and bis tosylate (pTSA Form 2). Each salt was treated withthe relevant solvent (10 vol, 30 mg in 300 μL) at 25° C. The sampleswere stirred for 10 minutes and then heated to 50° C. for 1 hour. Theresulting suspensions were matured for 24 hours and any resultingsolutions were allowed to evaporate slowly. After maturation orevaporation, the solids were filtered and air-dried before analysis byXRPD. Any solids that showed a new XRPD pattern were characterized by ₁HNMR, IC (if the counter-ion not seen in the NMR) and one weeks storageat 40° C. and 75% RH, if the amount of solid isolated was sufficient.

Tartrate.

Solvents used and results are shown below in Table 17.

TABLE 17 Observations Observations Ex. Solvent at 50° C. after treatmentXRPD P1 n-Heptane suspension suspension no change in form P2 Isopropylsuspension suspension no change in acetate form P3 MIBK suspensionsuspension no change in form P4 Acetone suspension suspension no changein form P5 2- gel gel TAR Form 3 Methoxyethanol P6 1,4-Dioxanesuspension suspension no change in form P7 Dichloromethane suspensionsuspension TAR Form 1 P8 Nitromethane gum gum TAR Form 1 P9 THF:Watergel suspension TAR Form 3 (95:5) P10 Acetone:Water gel suspension TARForm 3 (95:5)

From the 10 solvents used in this small polymorphism assessment, five ofthe tartrate samples changed in form. Samples from DCM and nitromethanechange to TAR Form 1, previously obtained in the initial saltexperiment. A new pattern, TAR Form 3, was found from 2-methoxyethanol,THF:water and acetone:water.

Limited characterisation was performed on the samples. These studiesindicated that the samples consistent with TAR Form 1 by XRPD convertedto TAR Form 3 on storage. By high resolution XRPD it was noted that thesample from 2-methoxyethanol displayed some differences to thediffractograms from the other two TAR Form 3 samples. It should be notedthat that samples which degraded on storage during the characterisationof scaled up tartrate salt (TAR Form 2) displayed peaks similar to TARForm 3. Owing to material availability, it was not possible to assessthe purity of the samples after the polymorphism assessment and storageconditions. Results are shown below in Table 18.

TABLE 18 High Res XRPD TAR Form 3 TAR Form 1 TAR Form 1 TAR Form 3 TARForm 3 ¹H-NMR Peak shifts to Peak shifts to Peak shifts Peak shifts Peakshifts (DMSO-d6) free base ~1 free base ~0.8 to free base ~0.8 to freebase ~1 to free base ~1 eq acid ~3 eq acid eq acid eq acid eq acid eqtrace DCM trace 2-methoxy- nitromethane ethanol Storage @ No change inChange in Change in No change No change 40° C./75% form form form inform in form RH TAR Form 3 TAR Form 3 TAR Form 3 TAR Form 3 TAR Form 3

Tosylate.

Solvents used and results are shown below in Table 19.

TABLE 19 Observations Observations after Ex. Solvent at 50° C. treatmentXRPD P11 n-Heptane suspension suspension no change in form P12 Isopropylsuspension suspension no change in acetate form P13 MIBK suspensionsuspension no change in form P14 Acetone suspension suspension no changein form P15 2- suspension suspension no change in Methoxyethanol formP16 1,4-Dioxane suspension suspension no change in form P17Dichloromethane suspension suspension no change in form P18 Nitromethanesuspension suspension no change in form P19 THF:Water solution gumamorphous (95:5) P20 Acetone:Water suspension suspension no change in(95:5) form

From the 10 solvents used, no change in crystalline form was noted in 9of the tosylate samples. The amorphous trace from the THF:water sampleis due to it becoming a gum after maturation. In this case, unlike theother solvents, the sample fully dissolved at 50° C. and a gumprecipitated out on cooling. This finding indicates this would not be agood solvent system for obtaining the crystalline tosylate salt. Whilstnone of the other samples changed form during this small polymorphismassessment, experiments showed that the tosylate salt had at least 2forms.

Oxalate.

Solvents used and results are shown below in Table 20.

TABLE 20 Observations Observations after Ex. Solvent at 50° C. treatmentXRPD P21 n-Heptane suspension lumpy solid no change in form P22Isopropyl suspension suspension no change in acetate form P23 MIBKsuspension suspension no change in form P24 Acetone suspensionsuspension no change in form P25 2- gel gel no change in Methoxyethanolform P26 1,4-Dioxane suspension suspension no change in form P27Dichloromethane suspension suspension no change in form P28 Nitromethanegel gel no change in form P29 THF:Water gel suspension OXA Form 2 (95:5)P30 Acetone:Water gel gel no change in (95:5) form

Out of the 10 solvents used in this small polymorphism assessment on theoxalate salt, only one change in form was observed. The sample fromTHF:water was found to be OXA Form 2, which was previously observed onexposure of OXA Form 1 and 3 to high humidity. It is believed that thisform is another hydrated form of the oxalate.

Conclusion.

The tartrate was found to change under both temperature and humidityconditions with a drop in purity noted under the latter conditions. Italso shows a propensity towards polymorphism with half of the samples inthe polymorphism assessment changing form. The tartrate is also a highlyhygroscopic material and displays complex thermal behaviour.

The oxalate changed form under humidity, but not temperature conditions.Only one of the samples from slurry experiments changed form. However,the material is hygroscopic and has undesirable thermal behaviour.

The tosylate salt, pTSA Form 2, did not change form under the majorityof conditions tested, including the polymorphism assessment slurries.The exception to this was in THF:water where an amorphous solid wasobtained. It is also the most crystalline salt identified during thecourse of the experiment and displays the best solid state properties.

Further Characterization of Tosylate Polymorphs.

First, the free base form of Example 1 was characterized in order toinvestigate its solid form and chemical properties. Consistent withpervious batches, the material was amorphous, with a purity of ca.97.6%. The material is not stable at elevated storage conditions(deliquesced at 25° C./97% RH and 40° C./75% RH). The ¹H-NMR isgenerally consistent with the structure and does not show anysignificant amounts of residual solvent aside from water. The amount ofwater was quantified by KF as 2.3%.

Preparation and Characterization of Amorphous Bis Tosylate Salt.

Initial conversion to the amorphous state increases the likelihood ofidentifying metastable forms in addition to the most thermodynamicallystable one. For this reason the amorphous bis tosylate was used for thisexperiment. Amorphous Example 1 was dissolved in DCM (1 g in 10 mL) atambient conditions. The resulting solution was treated withp-toluenesulfonic acid monohydrate (4044 μL, 1M in THF, 2.1 eq) atambient. The solution was fast evaporated on the rotary evaporator andthe recovered solid was analyzed by XRPD. The first such procedureyielded a semicrystalline pattern different from Form 2, which wasre-dissolved in DCM (10 mL), fast-evaporated and re-analyzed by XRPD andconfirmed to be Form 2. A second procedure described below yielded Form2. Amorphous Example 1 bis tosylate prepared as above was brieflycharacterized in order to investigate its solid form and chemicalproperties. XRPD analysis confirmed the material to be amorphous; ¹H-NMRwas consistent with structure and indicated 2 equivalents of tosylatecounterion; KF analysis indicated only 2.0% w/w water; and HPLC analysisindicated 97.6% purity, consistent with the starting material.

Preparation and Characterization of Form 2 Bis Tosylate Salt.

Example 1 was dissolved in ACN (2 g in 20 mL) at ambient conditions. Theresulting solution was stirred at 50° C. and treated withp-toluenesulfonic acid monohydrate (8088 μL, 1 M in THF, 2.1 eq). Afterstirring for 1 h at 50° C., the clear solution was cooled to −4° C.(0.1° C./min) and then remained at −4° C. overnight. The recovered solid(Form 2 Batch 1) was filtered under vacuum and air dried for 2 h anddried in the oven (RT, vacuum) overnight.

A second batch, described above (Form 2 Batch 2), was recovered afterattempting to make amorphous bis tosylate. A semicrystalline trace wasinitially observed by XRPD and DCM (20 vol) was added to try to dissolveit again. However, a suspension was formed. After 2 hours stirring at40° C., it was filtered, dried in the oven (RT/vacuum).

Both batches were consistent with Form 2 by XRPD. Thermal analyzesshowed an event corresponding to the loss of the water, followed by themelt of the sample at ca. 170° C. Both batches were also stable atelevated conditions for 1 week. These results correlate with Form 2obtained during previous experiments. During the PLM analysis of the twobatches, different morphologies were observed. Instead of theagglomerated particles observed to date, in the sample obtained byslurrying in DCM, needles were present. Although these were too small toanalyze by single crystal, this indicates there could be a possibilityof growing suitable crystals. Results are shown below in Table 21.

TABLE 21 Technique Form 2 Batch 1 Form 2 Batch 2 XRPD Form 2 Form 21H-NMR Consistent with Consistent with (DMSO-d6) structure + ca. 0.02 eqstructure No residual of CAN solvent KF 1.2% water 2.3% water HPLC(Purity %, 99.2 98.7 AUC) TGA 1.7% of weight loss 3.7% of weight lossbetween RT and 125° C. between RT and 85° C. Degradation fromDegradation from 280° C. 250° C. DSC Endotherm at 33.3° C. Endotherm at33.7° C. (58.2 J/g); Endotherm (77.9 J/g); Endotherm at 171.7° C. (22.8J/g) at 174.3° C. (52.2 J/g) Storage for 7 n/a Unchanged - Form 2 days @40° C./75% RH Storage for 7 n/a Unchanged - Form 2 days @ 25° C./97% RHPLM Irregular shape Needle shape particles particles up to 100 μm up to75 μm

Slurry Experiments.

Amorphous bis tosylate was suspended in the solvent system below (30 mgin 150 μl) at ambient conditions. The suspensions were shaken in thematuration chamber between ambient and 50° C. for 19 h (8 h cycles),then allowed to stand at ambient conditions for 10 min. An aliquot ofthe solid was air dried and analyzed by XRPD and then placed for 1 weekat 40° C./75% RH and re-analyzed. If solutions were recovered, thesolvents were allowed to evaporate at ambient conditions and theresidues were initially analyzed by XRPD. Results are shown below inTable 22.

TABLE 22 XRPD after 7 Appearance days @ Ex. Solvent after 19 h XRPD 40C. 75 RH P31 n-Heptane Suspension Amorphous Form 2 P32 Diethyl EtherSuspension Amorphous Form 2 P33 Ethyl Acetate Suspension Form 2 Form 2P34 Isopropyl Acetate Suspension Form 2 Form 2 P35 Methylisobutyl KetoneSuspension Crystalline Form 2 Form 3 P36 2-Propanol Suspension Form 2Form 2 P37 Methylethyl Ketone Suspension Crystalline Form 2 Form 5 P38Acetone Suspension Form 2 Form 2 P39 Dimethyl Sulfoxide Solution Oil n/aP40 Water Suspension Form 2 Form 2 P41 tert-Butylmethyl Ether SuspensionAmorphous Form 2 P42 Cyclohexane Suspension Amorphous Form 2 P431,4-Dioxane Suspension Crystalline Form 2 Form 4 P44 Toluene SuspensionSemi- Form 2 crystalline Form 3 P45 Chloroform Solution Form 2 Form 2P46 1,2-Dimethoxyethane Suspension Form 2 Form 2 P47 TetrahydrofuranSuspension Semi- Form 2 crystalline Form 3 P48 N,N- Solution Oil n/aDimethylformamide P49 Acetonitrile Suspension Form 2 Form 2 P50Nitromethane Suspension Form 2 Form 2 P51 N-Methylpyrrolidone SolutionOil n/a P52 Tetrahydrofuran:water Suspension Form 2 Form 2 95:5 P532-Propanol:water 95:5 Suspension Form 2 Form 2 P54 Acetone:water 95:5Suspension Form 2 Form 2

Solubility.

Crystalline bis tosylate Form 2 (30 mg) was treated with increasingvolumes of solvent (n-Heptane, diethyl ether, ethyl acetate, isopropylacetate, methylisobutyl ketone, 2-propanol, methylethyl ketone, acetone,dimethyl sulfoxide, water, tert-butylmethyl ether, cyclohexane,1,4-dioxane, toluene, chloroform, 1,2-dimethoxyethane, tetrahydrofuran,N,N-dimethylformamide, acetonitrile, nitromethane, N-methylpyrrolidone,tetrahydrofuran:water 95:5, 2-propanol:water 95:5, or acetone:water 95:5at 5, 10, 20, 30, 50, 70, or 100 vol total) until the material fullydissolved or until a maximum of 100 vol had been added. After eachaddition of solvent, the system was shaken gently for 10 minutes at 50°C. and then allowed to stand at ambient conditions for 5 minutes beforethe addition of a new aliquot of solvent. After the assessment wascompleted, any suspensions obtained were matured and clear solutionswere cooled to 4° C. and held isothermally. Example 1 bis tosylate Form2 remained a suspension up to in all solvents up to 100 vol except fordimethylsulfoxide and N,N-dimethylformamide, where it was a solution at5 vol, and tetrahydrofuran:water 95:5, where it was a solution at 10vol.

Maturation.

Suspensions obtained after the solubility assessment were shaken in thematuration chamber between ambient and 50° C. (8 h cycles). After 4 daysthe solids were filtered and air dried. Solids obtained were initiallyanalyzed by XRPD. Maturation in n-heptane, diethyl ether, ethyl acetate,isopropyl acetate, methylisobutyl ketone, 2-propanol, methylethylketone, acetone, tert-butylmethyl ether, cyclohexane, 1,4-dioxane,toluene, chloroform, 1,2-dimethoxyethane, tetrahydrofuran, acetonitrile,nitromethane, and 2-propanol:water 95:5 yielded filtered solids of Form2.

Slow Evaporation and Cooling.

Supernatants from the maturation experiments and solutions from coolingwere allowed to slowly evaporate at ambient conditions. The residueswere analyzed by XRPD. Slow evaporation from the following supernatantsyielded: dimethyl sulfoxide, oil; water, oil; N,N-dimethylformamide,gum; N-methylpyrrolidone, oil; tetrahydrofuran:water 95:5, gum; andacetone:water 95:5 gum. Solutions obtained after the solubilityassessment were placed in a fridge (4° C.) for 2 days and in a freezer(−20° C.) for another 2 days. No solids were recovered. Slow evaporationfrom two batches of n-heptane solution yielded a gum in from one batchand Form 2 from the other.

Slurring at 4° C. or 60° C.

Crystalline bis tosylate Form 2 (30 mg) was suspended in a solventsystem (10 vol, 3000. Suspensions were stirred at 4° C. or 60° C. overthe weekend. XRPD characterization of the 4° C. experiments yielded Form2 from ethyl acetate, 2-propanol, methylethyl ketone, tert-butylmethylether, N,N-dimethylformamide, acetonitrile, ethanol, and nitromethane, agum for tetrahydrofuran:water 95:5, and a solution forN-methylpyrrolidone. XRPD characterization of the 60° C. experimentsyielded Form 2 from ethyl acetate, 2-propanol, methylethyl ketone,tert-butylmethyl ether, acetone, 1,4-dioxane, acetonitrile, ethanol,nitromethane, and N-methylpyrrolidone.

Results.

The majority of samples isolated in the bis-tosylate polymorphexperiments were consistent with Form 2. However, three crystallinesolids (Form 3 (P35 from methylisobutyl ketone), Form 5 (P37 frommethylethyl ketone), and Form 4 (P43 from 1,4-dioxane)) displayed somedifferences. These were analyzed by high resolution X-ray powderdiffraction (HR-XRPD). Results are shown in FIG. #. Some extra peakswere noticed in Form 3 and Form 4. It was decided to characterize thesetwo new patterns further by HPLC, ¹H-NMR and DSC. Form 5 by highresolution was consistent with Form 2 however, when the initialdiffractogram (high throughput (HT) red trace) is compared with the newone (high resolution) a change can be seen. Based on these findings thematerial was concluded to be a metastable form, which rapidlytransformed to a more stable one, Form 2, by the time it was isolatedand analyzed by HR-XRPD.

Amorphous samples were matured for a further 7 days but no improvementin crystallinity was observed. The semicrystalline solids were alsomatured longer but did not show any improvement in crystallinity afterfurther 6 days cycling. As these (semicrystalline Form 3 fromtetrahydrofuran (P44) and semicrystalline Form 3 from toluene (P47))displayed some differences to Form 2 by XRPD they were furtherinvestigated by DSC, HPLC and ¹H-NMR. All solids obtained were alsostored for 7 days at 40° C. and 75% RH and re-analyzed by XRPD. Form 2was the only form observed after this time.

Form 3.

Thermal analysis of the solid from Form 3 (P35) showed an endothermicevent at 37.6° C. indicating that Form 3 may be a hydrate. A broadendotherm was also observed at 145.8° C. Analysis by VT-XRPD showed achange to something similar to Form 2 at 135° C. This correlates withthe diffractogram (data not shown) seen after 7 days at 40° C./75% RHindicating Form 3 is a metastable form, which readily converts to Form 2with heat and/or humidity.

Form 4.

¹H-NMR of Form 4 revealed two equivalents of 1,4-dioxane. This resultcorrelates with the endotherm at ca. 70.4° C. observed in the DSC. Afterthe desolvation, an endotherm at 169.3° C. was seen. This melt is thesame as Form 2, indicating Form 4 transforms to Form 2 once the solventis released. Due to the amount of material obtained TGA and KF analysiscould not be performed on the sample. Re-analysis by XRPD after 7 daysat 40° C./75% RH confirmed Form 4 is metastable form, which converts toForm 2 with heat and/or humidity. The HPLC purity of this form was alsolower than the others.

Analytical and Instrumental Methods

X-Ray Powder Diffraction (XRPD).

X-Ray Powder Diffraction patterns were collected on a Bruker AXS C2GADDS diffractometer using Cu Kα radiation (40 kV, 40 mA), automated XYZstage, laser video microscope for auto-sample positioning and a HiStar2-dimensional area detector. X-ray optics consists of a single Göbelmultilayer mirror coupled with a pinhole collimator of 0.3 mm. A weeklyperformance check is carried out using a certified standard NIST 1976Corundum (flat plate).

The beam divergence, i.e. the effective size of the X-ray beam on thesample, was approximately 4 mm. A θ-θ continuous scan mode was employedwith a sample—detector distance of 20 cm which gives an effective 20range of 3.2°-29.7°. Typically the sample would be exposed to the X-raybeam for 120 seconds. The software used for data collection was GADDSfor XP/2000 4.1.43 and the data were analyzed and presented usingDiffrac Plus EVA v15.0.0.0.

Samples run under ambient conditions were prepared as flat platespecimens using powder as received without grinding. Approximately 1-2mg of the sample was lightly pressed on a glass slide to obtain a flatsurface. Samples run under non-ambient conditions were mounted on asilicon wafer with heat-conducting compound. The sample was then heatedto the appropriate temperature at 20° C./min and subsequently heldisothermally for 1 minute before data collection was initiated.

High-Resolution XRPD.

X-Ray Powder Diffraction patterns were collected on a Bruker D8diffractometer using Cu Kα radiation (40 kV, 40 mA), θ-2θ goniometer,and divergence of V4 and receiving slits, a Ge monochromator and aLynxeye detector. The instrument is performance checked using acertified Corundum standard (NIST 1976). The software used for datacollection was Diffrac Plus XRD Commander v2.6.1 and the data wereanalyzed and presented using Diffrac Plus EVA v15.0.0.0.

Samples were run under ambient conditions as flat plate specimens usingpowder as received. The sample was gently packed into a cavity cut intopolished, zero-background (510) silicon wafer. The sample was rotated inits own plane during analysis. The details of the data collection are:angular range: 2 to 42° 2θ; step size: 0.05° 2θ; collection time: 0.5s/step.

Nuclear Magnetic Resonance (NMR).

NMR spectra were collected on a Bruker 400 MHz instrument equipped withan auto-sampler and controlled by a DRX400 console. Automatedexperiments were acquired using ICON-NMR v4.0.7 running with Topspinv1.3 using the standard Bruker loaded experiments. For non-routinespectroscopy, data were acquired through the use of Topspin alone.Samples were prepared in DMSO-d₆, unless otherwise stated. Off-lineanalysis was carried out using ACD Spectrus Processor 2012 or 2014.

Differential Scanning Calorimetry (DSC).

TA Instruments Q2000: DSC data were collected on a TA Instruments Q2000equipped with a 50 position auto-sampler. The calibration for thermalcapacity was carried out using sapphire and the calibration for energyand temperature was carried out using certified indium. Typically 0.5-3mg of each sample, in a pin-holed aluminium pan, was heated at 10°C./min from 25° C. to 300° C. A purge of dry nitrogen at 50 mL/min wasmaintained over the sample. Modulated temperature DSC was carried outusing an underlying heating rate of 2° C./min and temperature modulationparameters of ±0.318° C. (amplitude) every 60 seconds (period). Theinstrument control software was Advantage for Q Series v2.8.0.394 andThermal Advantage v5.5.3 and the data were analyzed using UniversalAnalysis v4.5A.

TA Instruments Discovery DSC: DSC data were collected on a TAInstruments Discovery DSC equipped with a 50 position auto-sampler. Thecalibration for thermal capacity was carried out using sapphire and thecalibration for energy and temperature was carried out using certifiedindium. Typically 0.5-3 mg of each sample, in a pin-holed aluminium pan,was heated at 10° C./min from 25° C. to 260° C.-300° C. A purge of drynitrogen at 50 mL/min was maintained over the sample. The instrumentcontrol and data analysis software was TRIOS v3.2.0.3877.

Thermo-Gravimetric Analysis (TGA). TGA data were collected on a TAInstruments Discovery TGA, equipped with a 25 position auto-sampler. Theinstrument was temperature calibrated using certified alumel and nickel.Typically 5-10 mg of each sample was loaded onto a pre-tared aluminiumDSC pan and heated at 10° C./min from ambient temperature to 350° C. Anitrogen purge at 25 mL/min was maintained over the sample. Theinstrument control and data analysis software was TRIOS v3.2.0.3877.

Polarised Light Microscopy (PLM).

Samples were studied on a Leica LM/DM polarised light microscope with adigital video camera for image capture. A small amount of each samplewas placed on a glass slide, mounted in immersion oil and covered with aglass slip, the individual particles being separated as well aspossible. The sample was viewed with appropriate magnification andpartially polarised light, coupled to a λ false-color filter.

Chemical Purity Determination by HPLC.

Purity analysis was performed on an Agilent HP1100 series systemequipped with a diode array detector and using ChemStation softwarevB.04.03 using one of the methods detailed below:

Parameter Value Type of method Reverse phase with gradient elutionSample Preparation 0.5 mg/mL in acetonitrile:water 1:1 Column SupelcoAscentis Express C18, 100 × 4.6 mm, 2.7 μm Column Temperature (° C.) 25Injection (μL) 5 Wavelength, Bandwidth 255, 90 (nm) Flow Rate (mL/min) 2Phase A 0.1% TFA in water Phase B 0.085% TFA in acetonitrile Time (min)% Phase A % Phase B Timetable 0 95 5 6 5 95 6.2 95 5 8 95 5

Parameter Value Type of method Reverse phase with gradient elutionColumn Phenomenex Luna, C18 (2) 5 μm 50 × 4.6 mm Column Temperature (°C.) Ambient Standard Injections (μL) 5 Wavelength, Bandwidth 260, 90(nm) Flow Rate (ml/min) 1 Phase A 0.1% TFA in water Phase B 0.085% TFAin acetonitrile Time (min) % Phase A % Phase B Timetable 0.0 95 5 5.0 955 12.0 75 25 20.0 75 25 30.0 40 60 35.0 40 60 40.0 5 95 40.1 95 5 45.195 5

Typically 5-20 mg of sample was placed in a tared mesh stainless steelbasket under ambient conditions. The sample was loaded and unloaded at40% RH and 25° C. (typical room conditions). A moisture sorptionisotherm was performed as outlined below (2 scans giving 1 completecycle). The standard isotherm was performed at 25° C. at 10% RHintervals over a 0-90% RH range. Data analysis was carried out usingMicrosoft Excel using DVS Analysis Suite v6.2 (or 6.1 or 6.0).

Gravimetric Vapour Sorption (GVS).

Sorption isotherms were obtained using a SMS DVS Intrinsic moisturesorption analyzer, controlled by DVS Intrinsic Control software v1.0.1.2(or v 1.0.1.3). The sample temperature was maintained at 25° C. by theinstrument controls. The humidity was controlled by mixing streams ofdry and wet nitrogen, with a total flow rate of 200 mL/min The relativehumidity was measured by a calibrated Rotronic probe (dynamic range of1.0-100% RH), located near the sample. The weight change, (massrelaxation) of the sample as a function of % RH was constantly monitoredby the microbalance (accuracy±0.005 mg).

Typically 5-20 mg of sample was placed in a tared mesh stainless steelbasket under ambient conditions. The sample was loaded and unloaded at40% RH and 25° C. (typical room conditions). A moisture sorptionisotherm was performed as outlined below (2 scans giving 1 completecycle). The standard isotherm was performed at 25° C. at 10% RHintervals over a 0-90% RH range. Data analysis was carried out usingMicrosoft Excel using DVS Analysis Suite v6.2 (or 6.1 or 6.0).

Parameter Value Adsorption - Scan 1 40-90 Desorption/Adsorption - Scan 290-0, 0-40 Intervals (% RH) 10 Number of Scans 2 Flow rate (ml/min) 200Temperature (° C.) 25 Stability (° C./min) 0.2 Sorption Time (hours) 6hour time out

Aqueous Solubility.

Aqueous solubility was determined by suspending sufficient compound inwater to give a maximum final concentration of ≥10 mg/ml of the parentfree-form of the compound. The suspension was equilibrated at 25° C. for24 hours then the pH was measured. The suspension was then filteredthrough a glass fibre C filter. The filtrate was then diluted by anappropriate factor. Quantitation was by HPLC with reference to astandard solution of approximately 0.2 mg/ml in DMSO. Different volumesof the standard, diluted and undiluted sample solutions were injected.The solubility was calculated using the peak areas determined byintegration of the peak found at the same retention time as theprincipal peak in the standard injection.

Parameter Value Type of method Reverse phase with gradient elutionColumn Phenomenex Luna, C18 (2) 5 μm 50 × 4.6 mm Column Temperature (°C.) 25 Standard Injections (μL) 1, 2, 3, 5, 7, 10 Test Injections (μL)1, 2, 3, 10, 15, 20 Detection: 260, 90 Wavelength, Bandwidth (nm) FlowRate (ml/min) 2 Phase A 0.1% TFA in water Phase B 0.085% TFA inacetonitrile Time (min) % Phase A % Phase B Timetable 0 95 5 1 80 20 2.35 95 3.3 5 95 3.5 95 5 4.4 95 5

Analysis was performed on an Agilent HP1100 series system equipped witha diode array detector and using ChemStation software vB.04.03.

Ion Chromatography (IC).

Data were collected on a Metrohm 930 Compact IC Flex with 858Professional autosampler and 800 Dosimo dosage unit monitor, using ICMagicNet software v3.1. Accurately weighed samples were prepared asstock solutions in an appropriate dissolving solution and dilutedappropriately prior to testing. Quantification was achieved bycomparison with standard solutions of known concentration of the ionbeing analyzed.

Type of method Anion exchange Cation exchange Column Metrosep A Supp5-150 Metrosep C 4 - 250 (4.0 × (4.0 × 150 mm) 250 mm) Column AmbientAmbient Temperature (° C.) Injection Various Various (μL) DetectionConductivity detector Conductivity detector Flow Rate 0.7 0.9 (ml/min)Eluent 3.2 mM sodium carbonate, 1.7 mM Nitric Acid 1.0 mM sodiumhydrogen 0.7 mM Dipicolinic acid in carbonate in a 5% acetone a 5%acetone aqueous aqueous solution. solution.

Maturation/Slurry Ripening.

Suspensions for maturation are placed in a platform shaker incubator(Heidolph Titramax/Inkubator 1000) and subjected to a series ofheat-cool cycles under shaking from ambient to 50° C. (8 hour cycles:heating to 50° C. for 4 hours and then the heating is switched off andthe sample gradually cools to ambient conditions for a further 4 hours).

Water Determination by Karl Fischer Titration (KF).

The water content of each sample was measured on a Metrohm 874 OvenSample Processor at 150° C. with 851 Titrano Coulometer using HydranalCoulomat AG oven reagent and nitrogen purge. Weighed solid samples wereintroduced into a sealed sample vial. Approx 10 mg of sample was usedper titration and duplicate determinations were made. Data collectionand analysis using Tiamo v2.2.

Biological Activity

The activity of the Examples above may be illustrated in the followingassays. Compounds listed above, which may not yet have been made and/ortested, are predicted to have activity in these assays.

Assaying the inhibition of KDM1A can be determined in vitro, in culturedcells, and in animals. There are a variety of spectrophotometric methodsto detect the results of demethylation of methylated lysines, viz.,detecting the products of KDM1A demethylase oxidative activity on apeptide fragment of at least 18 amino acid representing the N-terminusof the histone H3 substrate that contains a monomethyl at the fourthlysine residue. Hydrogen peroxide, one product of the KDM1A demethylasereaction, reacts with horseradish peroxidase and dihydroxyphenoxazine(ADHP) to produce the fluorescent compound resorufin (excitation=530-560nm:emission=590 nm). The KDM1A demethylase enzyme activity can obtainedfrom mammalian cells or tissues expressing KDM1A from an endogenous orrecombinant gene and purified or assayed from a whole cell extract.These methods can be used to determine the concentration of thedisclosed compounds can inhibit fifty percent of the enzyme activity(IC₅₀). In one aspect, the disclosed compounds exhibit inhibition fiftypercent of the KDM1A enzyme activity at a concentration of less than 500nM, less than 100 nM, less than 50 nM or less than 10 nM.

The association of KDM1A with other proteins can be determined by avariety of both in vitro and in vivo methods known to one skilled in theart. For example, the disruption of KDM1A with associated proteins canbe determined in an electromobility shift assay (EMSA). In variousaspects, the disruption of the physical association of KDM1A with CoRestby the disclosed compounds can be observed using EMSA. In anotherexample, the disruption of KDM1A with associated proteins can bedetermined by immunoprecipitation followed by separation of theco-precipitated proteins by mass spectroscopy or by get electrophoresis.In another example, the disruption of KDM1A association with CoRest canbe determined by the ability of KDM1A to act on a nucleosomal substratecontaining K4 or K9 methylated histone H3, a substrate that requires thepresence of both KDM1A and CoRest. The disclosed compounds could be usedto assay inhibition of CoRest association with KDM1A using nucleosomalsubstrate; such compounds may not inhibit KDM1A enzymatic activity asdetermined by the use of the histone H3 K4 methylated peptide substrate.

The inhibition of KDM1A can be determined in a cell-based assay. Forexample, KDM1A is an essential enzyme and prolonged inhibition of KDM1Awill result in cell death, thus cell growth inhibition, arrest of cellgrowth or cell death can be assayed. In another aspect, genes induced byandrogens and estrogens require KDM1A activity; inhibition by thedisclosed compounds of KDM1A will abrogate the induction of geneexpression in cells treated with androgens or estrogens. These effectscan be measured, e.g., using quantitative PCR of mRNA to measure themagnitude of gene expression for androgen- and estrogen-dependent genes.KDM1A activity is required for the repression of transcription ofspecific genes. Inhibition of KDM1A by the disclosed compounds couldde-repress the expression such genes in cell. These genes include Meis1,VEG-A, AIM1, HMOX1, VIM, SKAP1, BMP, EOMES, FOXA2, HNF4, SOX17, GH, PSA,pS2, GREB1, GR-1b, PRL, TSHB, SYN1, HBG, SCN1A, SCN2a, and SCN3A theexpression of which can be assayed using quantitative PCR of mRNA beforeand at various time following the treatment of cells with the disclosedcompounds. In another aspect, KDM1A is a regulator of leukemic stem cellpotential and is required for oncogenic transformation of myeloid cellsto acute myeloid leukemia (AML) by MLL-AF9. Inhibition of KDM1A inMLL-AF9-transformed cells grown in culture overcomes the arrest indifferentiation to resulting in a more mature cell expressing the CD11bsurface antigen, a monocytic cell antigen. Thus, inhibition of KDM1A canbe assayed using an AML cell line such as THP-1 grown in culturequantifying the proportion of cells newly expressing the CD11b antigenusing fluorescence activated cell sorting (FACS). A similar assay usingFACS to count cells displaying the CD14 or CD86 can be also used, eachof which are characteristic of more mature cells along themacrophage/monocytic lineage. Other cells lines derived from patientswith acute myeloid leukemia such as MV4;11 or MOLM-13 cells can be usedfor this assay. Other markers of differentiation along themacrophage/monocyte lineage can be similarly assayed by FACS such asCD14 and CD86. Other AML cell lines such as MPLM-13 or MV4;11 can beassayed for the induction of either specific genes mentioned above orthe differentiation markers as well as cell growth or apoptosis byAnnexin V staining and FACS enumeration.

The selectivity of the disclosed compounds for KDM1A can be determinedby assaying the IC₅₀ of the disclosed compounds for other FAD-dependentaminoxidases such as monoamine oxidase A (MAO-A), monoamine oxidase B(MAO-B), IL4I1, KDM1B, or SMOX. As such, a disclosed compound wouldinhibit KDM1A with an IC₅₀ that is 50-fold, or 100-fold or 250-fold or500-fold less than for MAO-A or MAO-B.

Additional Demethylase Assays

The histone demethylase assay can be performed essentially as describedin Shi, Y et al. Cell 199, 941-953 (2004). Briefly, bulk histones,histone peptides or nucleosomes are incubated with purified humanrecombinant KDM1A, in the histone demethylase activity (HDM) assaybuffer 1 (50 mM Tris pH 8.5, 50 mM KCl, 5 mM MgCl, 0.5% BSA, and 5%glycerol) from 30 minutes to 4 hours at 37° C. A typical reaction isconducted in 100 microliters in which either 20 micrograms of purifiedbulk histones or 3 micrograms of modified histone peptides are used assubstrates. Different amounts of KDM1A ranging from 1-20 micrograms areused in the reaction along with, as necessary, other co-factors such asFAD or CoREST, depending on the chosen substrate. The reaction mixtureis analyzed by SDS-PAGE and Western blotting using histonemethyl-specific antibodies or by formaldehyde formation assay to examinethe removal and conversion of the methyl group to formaldehyde, or bymass spectrometry in the case of peptide substrates to identify thedemethylated histone peptide.

Bulk histones (e.g., 4 mg) are incubated with the indicated amounts ofrecombinant proteins or complexes in histone demethylase (HDM) assaybuffer A (50 mM Tris pH8.5, 50 mM KCl, 5 mM MgCl, 5% glycerol, 0.2 mMphenylmethylsulphonyl fluoride and 1 mM dithiothreitol) in a finalvolume of 10 ml for 12-16 h at 37 8 C. For nucleosomes (0.3 mg) ormononucleosome (0.3 mg), HDM buffer A containing 0.1% NP40 can be used.The reaction mixture can then be analyzed by SDS-PAGE followed byWestern blotting. Antibodies against mono- or di-methyl K4 in histone H3and acetyl-K9/K14 of histone H3 are used to detect the degree ofmethylation and acetylation, respectively. Western blots are thenquantified by densitometry or by intensity of luminescence.

Alternatively, a standard fluorogenic assay can be used in which themethylated histone substrate is tethered to the bottom of a 96 wellplate (or to beads resting in the plate) using biotin conjugated to thehistone methylated substrate and strepavidin (SA) on beads or SAattached to the plate to secure the biotinylated substrate. Afterincubation of the KDM1A enzyme in histone demethylase buffer A, thedemethylated histone substrate can be detected using antibodies specificfor demethylated H3K4 substrate conjugated to a fluor or some otheragent that can be detected. A variation on that assay method wouldemploy an antibody directed against the methylated version of thehistone in which the amount of substrate is quantified before and afterincubation with the enzyme. Yet another version of a similar assay wouldemploy a fluorescence resonance energy transfer (FRET) system ofdetection in which the antibody recognizing the methylated version isconjugated or otherwise linked to an entity, e.g., a bead or a largecarrier molecule on which a fluorophore (donor) is attached and thefluorophore (acceptor) is bound to an entity linked to the substrate.

Alternatively, the production of H₂O₂ during the KDM1A reaction can bedetected fluometrically. In this system, the production of H₂O₂ isdetected in the HDM assay buffer after exposure to substrate, co-factorand enzyme using ADHP (10-Acetyl-3, 7-dihydroxyphenoxazine) as afluorogenic substrate for horse radish peroxidase (HRP). ADHP (alsoknown as Amplex Red Reagent) is the most stable and sensitivefluorogenic substrate for HRP. The florescent product is resorufin.Sensitivity can be as low as 10⁻¹⁵ M of target protein. The signal isread using a fluorescence microplate reader at excitation and emissionwavelengths of 530-560 nm and 590 nm, respectively.

Additionally, the KDM1A reaction can include other factors which mayinfluence the activity of KDM1A. Such factors might include CoREST, NuRDcomplexes, DNMT1, HDAC1, HDAC2, and HDAC3, for example, as proteinsknown to associate with KDM1A or KDM1A-containing complexes.Interactions that influence any aspect of the KDM1A activity includingspecificity for template, substrate, K_(m), K_(cat), or sensitivity toFAD concentrations can be assayed. For example, an in vitro interactionassay between KDM1A and CoREST can be performed adding recombinant KDM1A(e.g., 10 mg) and CoREST (e.g., 5 mg) mixed and incubated for 1 h at4-8° C., fractionated by Superdex 200 gel filtration column in a buffercontaining 20 mM Tris-HCl pH 7.9, 500 mM KCl, 10% glycerol, 0.2 mM EDTA,1 mM dithiothreitol, 0.1% Nonidet P40 and 0.2 mM phenylmethylsulphonylfluoride, and then analyzed by silver staining.

For co-immunoprecipitation of mononucleosomes with KDM1A and CoREST,nucleosomes (1.5 mg) can be digested with micrococcal nuclease andincubated with recombinant KDM1A (e.g., 1 mg), CoREST (e.g., 500 ng) orboth proteins in HDM buffer A containing 0.1% NP40 for 1 h at 4-8° C.Antibodies directed against KDM1A or CoREST attached to an affinityresin are added and after extensive washing with HDM buffer A containing0.1% NP40, the bound proteins are eluted with a wash buffer. KDM1Aactivity can be assayed in the eluate or the concentration of KDM1A canbe determined by quantitative Western blotting.

Compounds were tested in a 10-dose IC₅₀ mode fluorescence couplingenzyme assay with 3-fold serial dilution in duplicate starting at 100μM. The production of FAD-dependent H₂O₂ as a result of demethylaseactivity of LSD1 on 10 μM histone H3(1-21)K4me2 peptide substrate wasmeasured by coupling with HRP and Amplex Red to yield resorufin(fluorescence measured at Ex/Em=535/590 nm on EnVision, Perkin Elmer).Results are given below in Table 1.

TABLE 2 Biological Activity KDM1A Example IC₅₀ 1 28 2 13 3 37 4 12 5 5 622 7 5 8 5 9 6 10 34 11 129 12 17 13 6 14 27 15 50 16 98 17 13 18 10 193 20 5 21 45 22 23 5 24 19 25 22 26 11 27 65 28 24 29 20 30 4 31 13 32 833 2 34 ND 35 3 36 9 37 ND 38 8 41 11 42 18 43 20 44 10 45 20 46 13 4713 48 11 49 3 50 23 51 4 52 14 53 50 54 20 55 9 56 35 57 10 58 10 59 1760 ND 61 7 62 52 63 112 64 38 65 27 66 24 67 14 68 21 69 34 70 10 71 ND72 19 73 11 74 226 75 4 76 20 77 10 78 22 79 18 80 56 81 50 82 17 83 2884 15 85 8 86 7 89 5 90 6 91 210 92 4 93 19 94 12 95 18 96 8 97 5 98 2099 12 100 13 101 19 102 170 103 ND 104 9 105 13 106 10 107 5 108 5109 >1000 110 45 111 16 112 ND 113 23 114 ND 115 7 116 ND 117 20 118 13119 16 120 7 121 10 122 14 123 6 124 8 125 34 126 482 127 ND 128 2 12951 130 11 131 40 132 173 133 47 134 14 135 10 136 ND 137 ND 138 ND 139ND 140 ND 141 6 142 108 143 58 144 28 145 114 146 7 147 1 148 5 149 23150 24 151 11 152 3 153 13 154 ND 155 127 156 14 157 10 158 10 159 3 1604 161 46 162 6 163 35 164 10 165 157 166 14 167 11 168 11 169 64 170 65171 4 172 5 173 5 174 3 175 10 176 3 177 2 178 0.8 179 ND 180 2 181 4182 27 183 10 184 18 185 >1000 186 9 187 4 188 42Ex Vivo Differentiation of Purified Human CD34+ Cells into the ErythroidLineage

Human CD34+ cells isolated from the venous blood of healthy donors aftermobilization by granulocyte colony stimulating factor (G-CSF) are grownand differentiated ex vivo for a 14 day incubation using a two-phaseculture method described in Cui, S., et al. Mol Cell Biol 31, 3298-3311(2011). Cells are counted using a hemocytometer and viability determinedby trypan blue exclusion. Test article (candidate compounds) dissolvedin an appropriate solvent compatible with physiologic conditions isadded daily to fresh culture medium beginning on Day 4 through Day 14 ata range of test concentrations. Cell morphology and stage ofdifferentiation is determined by Wright-Giemsa staining.

Flow Cytometry to Determine Differentiation Surface Markers and HbFContent

Cultured erythroid cells are stained with phycoerythrin(PE)-Cy7-conjugated anti-CD34, PE-conjugated anti-CD71, andPECy5-conjugated anti-glycophorin A antibodies. To determine theconcentration of cytoplasmic HbF, cells are fixed in 0.05%glutaraldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 for5 minutes and stained with allophycocyanin-conjugated anti-HbF antibody.Stained cells are sorted and counted using a FACS analyzer.

Western Blots to Determine Presence and Concentration of KDM1A andHistone H3 and H3 Modifications.

Cells are lysed in Laemmli sample buffer and subjected to SDS-PAGE.Proteins are transferred from the gel to nitrocellulose and probed withantibodies against KDM1A, and/or histone H3, mono-methyl (H3K4me1)and/or dimethyl histone H3K4 (H3K4me2) and then probed withfluorescence-conjugated secondary antibodies. Proteins concentrationsare quantified with an imaging system.

Chromatin Immunoprecipitation (ChIP) Assays to Determine ProteinOccupancy at Genome-Specific Sites.

ChIP assays are carried out in an immunoprecipitation (IP) buffer withor without SDS depending on the sensitivity of the KDM1A antibody toSDS. Briefly, typically 3×107 cells are used per KMD1A ChIP and 3×106cells per H3K4me2 ChIP. After 10 minutes of 0.75% formaldehydetreatment, cells are harvested and sonicated in the ChIP lysis buffer(1% Triton X-100, 10 mM EDTA, 50 mM Tris-HCl and protease inhibitors) toproduce soluble chromatin with average sizes between 300 and 1000 bp.The chromatin samples are then diluted 10-fold in the dilution buffer (5mM EDTA, 25 mM Tris-HCl, 167 mM NaCl, and cocktails of proteaseinhibitors) and pre-cleaned for 1 hour using salmon sperm DNA/protein-Aagarose beads. Ten micrograms of rabbit anti-KDM1A antibody, 3microliters of anti-H3K4me2 or control antibodies are then added to eachsample and incubated overnight at 4° C. To collect the immunocomplexes,40 microliters of salmon sperm DNA/protein-A agarose beads are added tothe samples for 1 hour at 4° C. The beads are washed three times in washbuffer (0.1% Triton X-100, 5 mM EDTA, 30 mM Tris-HCl, 150 mM NaCl andthe washed once in wash buffer 2 (1% Triton X-100, 5 mM EDTA, 30 mMTris-HCl, 150 mM NaCl). The bound protein-DNA complexes are eluted with100 microliters of elution buffer (1% SDS, 0.1 M NaHCO3, 250 mM NaCl,and 0.2 micrograms protease K) and de-cross-linked at 65° C. for 4 hr.The de-crosslinked chromatin DNA is further purified by QIAquickpolymerase chain reaction (PCR) Purification Kit (Qiagen) and eluted in100 microliters of TE buffer. Four microliters of eluted DNA sample isused for each PCR reaction. Thirty-six PCR cycles can be used for KDM1AChIP and 32 PCR cycles for H3K4mme2 ChIP. Appropriate primers for lociof interest, e.g., the gamma globin gene, are used.

For globin-specific ChIP analysis, the assays are performed as describedin Cui, S., et al. Mol Cell Biol 31, 3298-3311 (2011). For example,ethylene glycol bis(succinimidyl succinate) or formaldehyde can be usedas a cross-linker. Antibodies against target proteins such as KDM1A andhistone H3 with or without methyl modifications can be used forimmunoprecipitation. DNA contained in the immunoprecipitate can bequantified by real-time quantitative PCR (RT-qPCR) assay using primerfor human embryonic, gamma, and adult beta-globin promoter sequences;primers for intergenic regions between the embryonic and gammaG-globingenes can be used as a negative control.

Hemoglobin Analysis by HPLC

Cells are lysed and can be analyzed for hemoglobin composition using theBio-Rad Variant II Hemoglobin Testing System equipped with anion-exchange HPLC column (Hercules).

Mouse Models for Testing Induction of Gamma Globin Gene Expression

Test article can be dissolved in a physiologically compatible solventfor injection into normal mice or mice transgenic for the yeastartificial chromosome (YAC) containing the entirety of the humanbeta-globin locus as described in Tanabe, O., et al. EMBO J 26,2295-2306 (2007) or portions of the human beta-globin locus. Testarticle can be administered daily intraperitoneally or subcutaneously orby gavage at appropriate test doses for up to 26 weeks. At intervals,peripheral whole blood and bone marrow cells are harvested to determinegene expression by RT-qPCR of the mouse embryonic beta-like globin genesor the beta-like globin composition of red cell lysates or in the casetransgenic mice carrying human beta-like globin genes both the human andmouse fetal γ- and adult β-globin genes.

Testing for Induction of Human Gamma Globin Gene Expression or HbF.

Patients with hemoglobinopathies including sickle cell disease andbeta-thalassemia might benefit from treatment with an inhibitor ofKDM1A. After appropriate dosing, the measure of HbF can be determined asdescribed above. Gamma globin gene expression can be assayed in bonemarrow cells using qPCR. Further, the clinical benefit of an agentinducing HbF can be measured as an increase in total hemoglobin, areduction in sickle cell crises, a decrease in transfusion dependence, adecrease in ineffective hematopoiesis, and decrease in inflammatorybiomarkers such as plasma levels of GDF15, etc.

Pharmacokinetics

The pharmacokinetic properties of the Examples above, includingabsorption, distribution, metabolism, and excretion, may be illustratedin the following assays. Compounds listed above, which may not yet havebeen made and/or tested, are predicted to have activity in these assays.

Metabolic Stability in Human and Murine Liver Microsomes

The metabolic stability of compounds disclosed herein in pooled humanliver microsomes (HLM) and pooled male mouse liver microsomes (MMLM) maybe determined according to the following protocol, in which theconcentrations of compounds in reaction systems were evaluated byLC/MS/MS for estimating the stability in liver microsomes.

Study Design

Pooled human liver microsomes (HMMCPL; PL050B) and pooled male mouseliver microsomes (MSMCPL; MS033) are purchased from CellzDirect(Invitrogen). Microsomes are stored at −80 C prior to use.

A master solution is prepared containing microsome (stock concentration5 mg/mL, volume 50 μL, final concentration 0.5 mg/mL), MgCl₂ solution(stock concentration 50 mM, volume 50 μL, final concentration 5 mM),phosphate buffer (stock concentration 200 mM, volume 250 μL, finalconcentration 100 mM), and water (volume 95 μL. Five μL of 200 μM testcompounds or control solution (control compound: verapamil) are thenadded. The final concentration of test compounds or verapamil in thereaction system is 2 μM. The mixture is pre-warmed at 37 C for 5 min.

The reaction is started with the addition of 50 μL of 10 mM NADPHsolution at the final concentration of 1 mM and carried out at 37 C. 50μL of ultra-pure H₂O is used instead of NADPH solution in the negativecontrol.

Aliquots of 50 μL are taken from the reaction solution at 0 and 30 min.The reaction is stopped by the addition of 3 volumes of cold methanolwith IS (200 nM imipramine, 200 nM labetalol and 2 μM ketoprofen) at thedesignated time points. Samples are centrifuged at 16,000 g for 10minutes to precipitate protein. Aliquot of 100 μL of the supernatant isdiluted by 100 μL ultra-pure H₂O, and the mixture is used for LC/MS/MSanalysis. All experiments may be performed in duplicate.

Bioanalytical Method

Samples are analyzed using liquid chromatography-mass spectrometry. AnLC system may comprise, for example, a Shimadzu liquid chromatographseparation system equipped with degasser DGU-20A3, solvent delivery unitLC-20AD, system controller CBM-20A, column oven CTO-10ASVP and CTCAnalytics HTC PAL System. Chromatographic conditions may include aPhenomenex column, 5.0μ C18 (2.0×50 mm); a mobile phase of 0.1% formicacid in acetonitrile and 0.1% formic acid in water; an elution rate of500 μL/min; column temperature 25 C; injection volume 10 μL. Massspectrometric analysis is performed using an API 4000 instrument from ABInc. (Canada) with an ESI interface. Data acquisition and control systemare created using, e.g., Analyst 1.5.1 software from ABI Inc. A turbospray ion source and electrospray ionization are employed in a multiplereaction monitoring (MRM) scan. Additional parameters include: collisiongas, 6 L/min; curtain gas, 30 L/min; nebulize gas, 50 L/min; auxiliarygas, 50 L/min; temperature, 500 C; ionspray voltage, +5500 v (positiveMRM). Quadripoles Q1 and Q3 are set to 456.2 and 200.2, respectively;declustering potential (DP), entrance potential (EP), and collision cellentrance potential (CE) are set to 120, 10, and 55 v, respectively;collision cell exit potential (CXP) is 12 v.

Analysis

All calculations may be carried out using Microsoft Excel. Peak areasare determined from extracted ion chromatograms. The control compoundsare included in the assay. Any value of the compounds that is not withinthe specified limits is rejected and the experiment is repeated. Thereaction system without the cofactors is used to exclude any misleadingfactor that results from instability of chemical itself.

Results

In pharmaceutical and medicinal chemistry, it is often desirable to havepotent compounds with properties lending suitability to drugdevelopment, such as solubility, stability, and reliability ofsynthesis. For example, compounds comprising a moiety—(CH₂)_(m)—Y—(CH₂)_(n)—Z—resulting in —(CH₂)₃—NR^(4b)— have shown to beamenable to synthesis with few undesired by products and/or relativelygood stability. Additionally, as disclosed in the table above, compoundswherein R³ is aryl substituted with heteroaryl, cyano, or —S(O)₂N(CH₃)₂(called R⁶ or R^(6a)) have in general demonstrated good potency.

Compositions

The following are examples of compositions which may be used to delivercompounds disclosed herein. These may be encapsulated or wet granulatedusing methods known in the art.

Composition Example 1

Ingredients Concentration (w/w %) Compound of Formula I 30% Lactose 68%Magnesium Stearate  2%

Composition Example 2

Ingredients Concentration (w/w %) Compound of Formula I 50% Lactose 48%Magnesium Stearate  2%

From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions.

Other Embodiments

The detailed description set-forth above is provided to aid thoseskilled in the art in practicing the present disclosure. However, thedisclosure described and claimed herein is not to be limited in scope bythe specific embodiments herein disclosed because these embodiments areintended as illustration of several aspects of the disclosure. Anyequivalent embodiments are intended to be within the scope of thisdisclosure. Indeed, various modifications of the disclosure in additionto those shown and described herein will become apparent to thoseskilled in the art from the foregoing description, which do not departfrom the spirit or scope of the present inventive discovery. Suchmodifications are also intended to fall within the scope of the appendedclaims.

All references cited in this specification are hereby incorporated byreference. The discussion of the references herein is intended merely tosummarize the assertions made by their authors and no admission is madethat any reference constitutes prior art relevant to patentability.Applicant reserves the right to challenge the accuracy and pertinence ofthe cited references.

What is claimed is:
 1. The compound

or a salt thereof.
 2. A salt of the formula:

wherein: X is chosen from tosylate, sulfate, tartrate, oxalate,besylate, fumarate, citric, esylate, and malate; and q is an integerchosen from 1 and
 2. 3. The salt as recited in claim 2, wherein X istosylate.
 4. The salt as recited in claim 2, wherein q is
 2. 5.N—((S)-5-((1R,2S)-2-(4-fluorophenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl)-4-(1H-1,2,3-triazol-1-yl)benzamidebis-tosylate.
 6. A tosylate salt of a compound as recited in claim
 1. 7.A pharmaceutical composition comprising a compound as recited in claim1, or a salt thereof, together with a pharmaceutically acceptablecarrier.
 8. The pharmaceutical composition as recited in claim 7,formulated for oral administration.
 9. The pharmaceutical composition asrecited in claim 8, additionally comprising another therapeutic agent.